Immunoprecipitation (IP) is a common research technique used to precipitate a specific protein (or protein complex) of interest out of a heterogeneous lysate. The concept of precipitating proteins out of solution is often called a "pull-down". There are a variety of methods to achieve immunoprecipitation, the most common of which uses antibodies bound to magnetic or agarose beads. This process can be used to isolate and concentrate a particular protein or complex from a sample containing many thousands of different proteins, and is often used to study protein-protein interactions through Co-IP experiments.
Novus offers a range of support options for IP when you buy a product to help you with your IP experiment including:
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB400-148SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NB100-56726 | |||
| BSA Free | NB100-56722SS | |||
| Azide and BSA Free | NB100-56726SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB110-58851SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-74555-0.025ml | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-42225SS | |||
| Azide and BSA Free | NBP2-80654 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-24737SS | |||
| Azide and BSA Free | NBP2-33259 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | FAB3567P-100 | |||
| Carrier Free | FAB3567P-025 | |||
| Carrier Free | FAB3567F-100 | |||
| Carrier Free | FAB3567F-025 | |||
| Carrier Free | FAB3567A-100 | |||
| Carrier Free | FAB3567A-025 | |||
| Carrier Free | FAB3567N-100 | |||
| Carrier Free | FAB3567N-025 | |||
| Carrier Free | FAB3567C-100 | |||
| Carrier Free | FAB3567C-025 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-33006-0.2mg | |||
| BSA Free | NBP2-33006-0.02mg | |||
| Azide and BSA Free | NBP2-34522-0.2mg | |||
| Azide and BSA Free | NBP2-34522-0.1mg | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB110-93479SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-139SS | |||
![Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - NPC proteins in mouse lungs. Western blot of wild type (W) littermates, NPC1 (Mut) or NPC2 (Mut) mutant mouse lungs using anti-NPC1 or -NPC2 antibody. beta-actin used as a loading control. 30 ug protein/lane. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0067084), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Western-Blot-NB400-148-img0015.jpg "Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - NPC proteins in mouse lungs. Western blot of wild type (W) littermates, NPC1 (Mut) or NPC2 (Mut) mutant mouse lungs using anti-NPC1 or -NPC2 antibody. beta-actin used as a loading control. 30 ug protein/lane. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0067084), licensed under a CC-BY license.")
![Immunocytochemistry/Immunofluorescence: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - NPC1 antibody was tested in HeLa cells with DyLight 488 (green). Nuclei were counterstained with DAPI (blue).](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB400-148-img0010.jpg)
![Immunohistochemistry-Paraffin: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Staining of human brain, cortex, neurons and astrocytes.](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Immunohistochemistry-Paraffin-NB400-148-img0011.jpg)
![Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Protein expression analysis of normal and NPC-deficient cells after HIV-1 infection. Cells were uninfected or infected with VSVG-HIV-1 and harvested 96 h post-infection. NPC2, NPC1, and beta-actin protein expression was detected via Western blotting in uninfected and infected cells. Gag expression was also detected in the infected cells. Image collected and cropped by CiteAb from the following publication (//virologyj.biomedcentral.com/articles/10.1186/1743-422X-9-31), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Western-Blot-NB400-148-img0016.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Immunocytochemical staining with corresponding colocalization analysis of ALIX (green) and NPC-1 (red) in fibroblasts exposed to LLOMe for 15 min. Nuclei are stained with DAPI (blue) and merged images show colocalization in yellow (n = 5). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32409651/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Niemann-Pick C1 Antibody - BSA Free-Immunocytochemistry-Immunofluorescence-NB400-148-img0017.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and NPC1 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:1000 of Rabbit Anti-Human NPC1 Polyclonal Antibody (Catalog # NB400-148) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for NPC1 at approximately 240-260 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Knockout-Validated-NB400-148-img0014.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - HeLa lysate, BHK lysate, A431 lysate tested at 1:1000. Image from verified customer review.](http://images.novusbio.com/fullsize/Niemann-Pick-C1-Antibody---BSA-Free-Knockdown-Validated-NB400-148-img0012.jpg)
![Immunocytochemistry/ Immunofluorescence: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - ESCRT complex recruited to LLOMe-damaged lysosomes promotes cell survival.Human fibroblasts were exposed to 1 mM LLOMe & when indicated, the cell permeable calcium chelator BAPTA-AM (1 µM) was added 10 min prior to LLOMe exposure. Immunocytochemical staining with corresponding colocalization analysis of a CHMP4B (green) & LAMP2 (red), b ALIX (green) & NPC-1 (red), & c CHMP4B (green) & LAMP2 (red) in fibroblasts exposed to LLOMe for 15 min. Nuclei are stained with DAPI (blue) & merged images show colocalization in yellow (n = 5). d Caspase-3 like activity in cells exposed to LLOMe for 8 h (n = 3). e Quantification of apoptotic & necrotic cells using Annexin V/PI staining in fibroblasts exposed to LLOMe for 6 h (n = 3). f Immunoblot of LAMP2 in digitonin-extracted cytosolic fractions from cells exposed to LLOMe for 2 h (n = 8) with corresponding quantitative analysis. *p < 0.05, **p < 0.01 & ***p < 0.001 compared to control (a–b) or LLOMe only (c–f). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32409651), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb400-148_rabbit-polyclonal-niemann-pick-c1-antibody-310202416165630.jpg)
![Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Expression of proteins involved in extracellular lipid uptake & intracellular cholesterol trafficking following JQ1 (0.4 µM) administration to HepG2 cells for 48 hours. (A) Representative Western blot (left panel) & densitometric analysis (right panel) of SR-B1. n = 6 independent experiments. (B) Immunofluorescence analysis of SR-B1 (green). Nuclei were counterstained with DAPI. n = 3 different experiments. Scale bar: 50 µm. (C) Representative Western blot (left panel) & densitometric analysis (right panel) of LDLr. n = 5 independent experiments. (D) LDLr immunofluorescence (green). Nuclei were counterstained with DAPI. n = 3 different experiments. (E–F) Representative Western blots & densitometric analysis of NPC1 & TMEM97. Tubulin was chosen as loading control. n = 6 independent experiments. Data represent means ± SD. Statistical analysis was performed by using unpaired Student’s t test. ** p < 0.01; *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32075110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb400-148_rabbit-polyclonal-niemann-pick-c1-antibody-310202416163786.jpg)
![Western Blot: Niemann-Pick C1 Antibody - BSA Free [NB400-148] - Effects of p75NTR modulation by LM11A-31 on cholesterol metabolism & secretion. (A–D) Representative Western blot & densitometric analysis of HMGCR, ABCA1, NPC1, & ApoE in U373 cells treated with vehicle (Ctrl) & LM11A-31 (0.1 µM) for 48 h. n = 4 different experiments. Vinculin served as a housekeeping protein to normalize protein loading. (E) Quantification of ApoE amount (µg/mL) by ELISA assay in culture medium from vehicle- & LM11A-31-treated U373 cells. n = 3 different experiments. (F) Cholesterol quantification (total cholesterol, free cholesterol, & cholesteryl esters) in the culture medium of U373 cells treated with vehicle (Ctrl) & LM11A-31 (0.1 µM) for 48 h. n = 3 different experiments. Data represent means ± SD. Statistical analysis was carried out by using unpaired Student’s t test. *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35563230), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb400-148_rabbit-polyclonal-niemann-pick-c1-antibody-31020241616058.jpg)
![Flow Cytometry: TLR2 Antibody (TL2.1) [NB100-56722] - Analysis using the FITC conjugate of NB100-56726. Surface staining of stable HEK293/hTLR2 cells (IML-202, red) and vector control cells (IML-200, green) using TLR2 antibody at 1 ug/10^6 cells.](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Flow-Cytometry-NB100-56722-img0005.jpg "Flow Cytometry: TLR2 Antibody (TL2.1) [NB100-56722] - Analysis using the FITC conjugate of NB100-56726. Surface staining of stable HEK293/hTLR2 cells (IML-202, red) and vector control cells (IML-200, green) using TLR2 antibody at 1 ug/10^6 cells.")
![Western Blot: TLR2 Antibody (TL2.1) [NB100-56722] - Expression of TLR2 was determined by western blotting. The immunoreactivity of the anti-TLR2 was confirmed with human intestinal lysate. Data are representative for analysis of >=2 independent experiments. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0060671) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Western-Blot-NB100-56722-img0015.jpg)
![Western Blot: TLR2 Antibody (TL2.1) [NB100-56722] - Total protein from human THP-1 and U937 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-TLR2 (NB100-56722) in 5% BSA-TBST and detected with an anti-mouse HRP secondary antibody using NovaLume chemiluminescence detection reagent (NPB2-61915).](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Western-Blot-NB100-56722-img0016.jpg)
![Flow Cytometry: TLR2 Antibody (TL2.1) [NB100-56722] - Monitoring of TLR2 expression in normal B cells and primary MCL cells cultured for 24h in the absence/presence of CD40L, IL-4, and TLR ligands. Filled curves refer to isotypic control, empty curves to the TLR2 antibody (NB100-56722) of untreated samples. Dotted histograms represent CD40L+IL4 activated cells, dashed curves indicate CD40L+IL4 activated cells further treated with TLR ligands. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0153823) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Flow-Cytometry-NB100-56722-img0013.jpg)
![Flow (Cell Surface): TLR2 Antibody (TL2.1) [NB100-56722] - Flow (Cell Surface): - Azide Free [NB100-56726] - Analysis using the PE conjugate of NB100-56726. Staining of TLR2 on stable transfected cell line using this antibody at 1 ug/10^6 cells. Green: isotype control. Red: anti-TLR2 antibody. Image using the Azide Free form of this antibody.](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Flow-Cell-Surface-NB100-56722-img0010.jpg)
![Block/Neutralize: TLR2 Antibody (TL2.1) [NB100-56722] - Human monocyte-derived DCs were pre-incubated wit TLR-neutralizing antibodies including TLR2 antibody (NB100-56722) for 1 h. Afterwards, DCs were infected with H. pylori G27 (MOI 5) and the levels of the costimulatory molecule CD86 and the maturation marker CD83 were analyzed by FACS on CD11c+ cells 24 h post-infection. Data are presented as mean +/- SD of five independent experiments. *p<=0.05, **p<=0.005, ***p<=0.0005. Asterisks on top of bars indicate significance relative to non-neutralized, H. pylori-primed control cells. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0104804) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/TLR2-Antibody-TL2-1-Block-Neutralize-NB100-56722-img0014.jpg)
![Flow Cytometry: TLR2 Antibody (TL2.1) - BSA Free [NB100-56722] - Intracellular flow analysis of TLR2 in 1x10^6 PBMCs using 2 ug of NB100-56722. Shaded histogram represents cells without antibody; green represents isotype control; purple represents anti-TLR2 antibody.](http://images.novusbio.com/fullsize/nb100-56722_mouse-monoclonal-tlr2-antibody-tl2-1-17720241003212.jpg)
![Western Blot: Nox4 Antibody [NB110-58851] - Whole cell protein from human HeLa, Hek293, HepG2, mouse Neuro2A and rat PC12 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% BSA in TBST. The membrane was probed with 2.0 ug/ml anti-Nox4 in 1% BSA and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Nox4 is detected at approx. 70 kDa (arrowhead)](http://images.novusbio.com/fullsize/Nox4-Antibody-Western-Blot-NB110-58851-img0015.jpg "Western Blot: Nox4 Antibody [NB110-58851] - Whole cell protein from human HeLa, Hek293, HepG2, mouse Neuro2A and rat PC12 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% BSA in TBST. The membrane was probed with 2.0 ug/ml anti-Nox4 in 1% BSA and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Nox4 is detected at approx. 70 kDa (arrowhead)")
![Immunocytochemistry/Immunofluorescence: Nox4 Antibody [NB110-58851] - HeLa cells were fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% Triton X-100 in PBS for 10 min. Cells were incubated with antibodies to Nox4 (NB110-58851) and the ER marker Calreticulin (NBP1-47518), each at 20 ug/ml for 1 hour at room temperature. The coverslips were washed 3x in PBS and incubated with Alexa-Fluor 488 anti-rabbit secondary antibody. (green) and DyLight 550 anti-mouse secondary antibody. The merged image shows the co-localization of Nox4 and Calreticulin in the ER.](http://images.novusbio.com/fullsize/Nox4-Antibody-Immunocytochemistry-Immunofluorescence-NB110-58851-img0012.jpg)
![Immunohistochemistry: Nox4 Antibody [NB110-58851] - The expression of Nox4, p21, and Rb was upregulated in degenerative discs. Immunohistochemical staining for Nox4 in NP specimens. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/omcl/2017/7426458/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nox4-Antibody-Immunohistochemistry-NB110-58851-img0019.jpg)
![Immunohistochemistry: Nox4 Antibody [NB110-58851] - Analysis using the Biotin conjugate of NB110-58851. Staining of NOX4 in proximal convoluted tubules of human kidney.](http://images.novusbio.com/fullsize/Nox4-Antibody-Immunohistochemistry-NB110-58851-img0005.jpg)
![Immunocytochemistry/Immunofluorescence: Nox4 Antibody [NB110-58851] - HeLa cells were fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% triton X-100 in PBS for 10 min. Cells were incubated with NB110-58851 at 20 ug/ml for 1 hour at room temperature, washed 3x in PBS and incubated with Alexa-Fluor 488 anti-rabbit secondary antibody. Nox4 (Green) was detected in the ER.](http://images.novusbio.com/fullsize/Nox4-Antibody-Immunocytochemistry-Immunofluorescence-NB110-58851-img0013.jpg)
![Western Blot: Nox4 Antibody [NB110-58851] - Alternative splicing of NOX4 in rat kidney and heart. Alternative splicing of NOX4 in human hearts. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2017.00935/full), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nox4-Antibody-Western-Blot-NB110-58851-img0020.jpg)
![Western Blot: Nox4 Antibody [NB110-58851] - Analysis using the Biotin conjugate of NB110-58851. Detection of NOX4 in human kidney lysates uisng NB110-58851 at 0.5 ug/ml. Band observed at 31 kDa may represent reported splice isoform.](http://images.novusbio.com/fullsize/Nox4-Antibody-Western-Blot-NB110-58851-img0016.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: Nox4 Antibody - BSA Free [NB110-58851] - Nox4 Antibody [NB110-58851] - Reduced levels of NADPH oxidases 1 and 4 at 18% versus 5% O2. Representative Western blots showing Nox1 and beta-tubulin in PC-3 cells or Nox4 and beta-tubulin in C2C12 cells, at 5% and 18% O2. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/omcl/2018/8238459/), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nox4-Antibody-Western-Blot-NB110-58851-img0021.jpg)
![Western Blot: Nox4 Antibody - BSA Free [NB110-58851] - Small interfering RNA against Nox4 (siNox4) alleviated oxidative stress & DNA damage induced by high oxygen tension in NP cells. (a, b) RT-qPCR analysis (N = 3) & representative immunoblot analysis of Nox4 in NP cells. The knockdown of Nox4 in NP cells was confirmed. (c) ROS production in NP cells (N = 3). (d) RT-qPCR analysis of MsrbA, MsrB1, & MsrB2 in NP cells (N = 3). (e, f) Immunofluorescence staining of gamma -H2A.X & percentage of gamma -H2A.X-positive cells in NP cells (N = 6). NP cells were transfected with siNox4 or scrambled siRNA control (siCtrl) before high oxygen tension treatment. ∗, P value < 0.05, error bars represent standard error. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29147462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb110-58851_rabbit-polyclonal-nox4-antibody-310202415395922.jpg)
![Western Blot: Nox4 Antibody - BSA Free [NB110-58851] - Nox4 overexpression boosted ROS production & induced DNA damage in NP cells. (a) RT-qPCR analysis (N = 4) of Nox4, MsrB1, & MsrB2 in NP cells overexpressing Nox4. (b) Representative immunoblot analysis of Nox4 in NP cells overexpressing Nox4. (c) ROS production in NP cells overexpressing Nox4 (N = 3). (d, e) Immunofluorescence staining of gamma -H2A.X & percentage of gamma -H2A.X-positive cells in NP cells overexpressing Nox4 (N = 8). NP cells were transfected with Nox4 vectors for Nox4 overexpression. ∗, P value < 0.05, error bars represent standard error. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29147462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb110-58851_rabbit-polyclonal-nox4-antibody-310202415171335.jpg)
![Western Blot: Nox4 Antibody - BSA Free [NB110-58851] - Alternative splicing of NOX4 in rat kidney & heart (A). Alternative splicing of NOX4 in human hearts (B). Quantitative evaluation of spliced NOX4 isoforms in ICM samples (C). Quantitative evaluation of spliced NOX4 isoforms in DCM samples (D). Data are mean ± S.E.M. n = 5/group. *p < 0.05. LV, left ventricle; IVS, interventricular septum; RV, right ventricle; ICM, ischemic cardiomyopathy; DCM, dilated cardiomyopathy. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29204124), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb110-58851_rabbit-polyclonal-nox4-antibody-310202415371935.jpg)
![Immunohistochemistry-Paraffin: Tryptophan hydroxylase 2 Antibody [NB100-74555] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse brain using Tryptophan hydroxylase 2 antibody at 1:25 dilution. The staining was developed using HRP-DAB detection method and the nuclei were counterstained with hematoxylin. The antibody generated staining of the neurons.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Immunohistochemistry-Paraffin-NB100-74555-img0009.jpg "Immunohistochemistry-Paraffin: Tryptophan hydroxylase 2 Antibody [NB100-74555] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse brain using Tryptophan hydroxylase 2 antibody at 1:25 dilution. The staining was developed using HRP-DAB detection method and the nuclei were counterstained with hematoxylin. The antibody generated staining of the neurons.")
![Western Blot: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Total protein from mouse brain, stomach and human brain was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Tryptophan Hydroxylase 2 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Western-Blot-NB100-74555-img0010.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Tryptophan hydroxylase 2 Antibody [NB100-74555] - The 5-HT and Tryptophan hydroxylase 2 in the raphe nuclei of the brain. The 5-HT- and Tryptophan hydroxylase 2-positive double stained cells in the dorsal raphe nuclei. Representative photomicrographs were taken at magnifications of 100x. Quantification of 5-HT and Tryptophan hydroxylase 2. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2218-273X/10/1/71), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-74555-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Analysis in mouse brain cells. Image courtesy of product review by Assaf Vestin.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-74555-img0006.jpg)
![Immunohistochemistry Free-Floating: Tryptophan hydroxylase 2 Antibody [NB100-74555] - PFA-perfused mouse midbrain serotoninergic neurons. AlexaFluor488 used as secondary antibody. Neurons pseudocolored in yellow hot LUT. Image from verified customer review.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Immunohistochemistry-Free-Floating-NB100-74555-img0008.jpg)
![Immunoprecipitation: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Analysis of raphe nuclei isolated from Sprague-Dawley rat brains using TPH2 Polyclonal Antibody. PA1-778 was coupled to magnetic beads and added to raphe nuclei whole cell extracts for 15 minutes at RT. Immunoprecipitated proteins were assessed by Western blotting (WB), using either mouse anti-TPH2 (Ms TPH2, Lane 2) or sheep anti-TPH2 antibodies (Sh TPH2, Lane 3). As a positive control for the Western blot, non-immunoprecipitated brain lysates were probed with the mouse anti-TPH2 antibody (Lane 1). As a specificity control, a blot containing immunoprecipitated proteins was probed with an antibody specific for Tyrosine Hydroxylase (TH, Lane 4). Data courtesy of the Innovators Program.](http://images.novusbio.com/fullsize/Tryptophan-hydroxylase-2-Antibody-Immunoprecipitation-NB100-74555-img0007.jpg)
![Western Blot: CD63 Antibody (H5C6) [NBP2-42225] - THP1 whole cell protein was separated by SDS-PAGE on a 12% gel and transferred to PVDF membrane. The membrane was probed with anti-CD63 antibody at 2 ug/mL and detected with an anti-mouse HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Western-Blot-NBP2-42225-img0002.jpg "Western Blot: CD63 Antibody (H5C6) [NBP2-42225] - THP1 whole cell protein was separated by SDS-PAGE on a 12% gel and transferred to PVDF membrane. The membrane was probed with anti-CD63 antibody at 2 ug/mL and detected with an anti-mouse HRP secondary antibody using chemiluminescence.")
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - The CD63 (H5C6) antibody was tested in HeLa cells at a 1:50 dilution against DyLight 488 (Green). Actin and nuclei were counterstained against Phalloidin 568 (Red) and DAPI (Blue), respectively.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0001.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD63 Antibody [H5C6] conjugated to DyLight 550 (NBP2-42225R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0014.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on SK-MEL-28 cells with CD63 (H5C6) Antibody NBP2-42225R (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0013.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - MDCK cells stained with CD63 antibody at a dilution of 1:50 followed by Donkey anti-mouse secondary antibody conjugated with Alexa Fluor 488 (1:500). Nuclei were stained with Hoechst 33342. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0006.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - U2OS cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD63 Antibody [H5C6] conjugated to Alexa Fluor 488 (NBP2-42225AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0015.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD63 Antibody [H5C6] conjugated to Biotin (NBP2-42225B) at 5 ug/ml for 60 minutes at room temperature and detected with Streptavidin Protein conjugated to DyLight 488 at a 1:500 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD63 Antibody [H5C6] conjugated to Biotin (NBP2-42225B) at 5 ug/ml for 1 hour at room temperature and detected with Streptavidin conjugated to DyLight 488. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0016.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-CD63 [H5C6] conjugated to Alexa Fluor 488 [NBP2-42225AF488] at 10ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0008.jpg)
![Western Blot: CD63 Antibody (H5C6) - BSA Free [NBP2-42225] - Analysis of common EV markers, where 10 ug of total protein was loaded in each lane. cASCs-EVs expressed CD63 and CD9, while beta actin and lamin A showed lower expression. The displayed data represent at least three repeated experiments with consistent results. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32040478/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-BSA-Free-Western-Blot-NBP2-42225-img0019.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on HeLa cells with CD63 [H5C6] Antibody NBP2-42225AF594 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 594.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0018.jpg)
![Western Blot: CD63 Antibody (H5C6) [NBP2-42225] - The same samples and volumes were run under reducing and non-reducing conditions. All procedures were performed in parallel for both conditions. L- ladder +- exosomes. Image visualized with HRP linked to secondary antibody. Western blot image submitted by a verified customer review.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Western-Blot-NBP2-42225-img0007.jpg)
![Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [NBP2-42225] - HEK cells stained with CD63 antibody at a dilution of 1:50 followed by Donkey anti-mouse secondary antibody conjugated with Alexa Fluor 488 (1:500). Nuclei were stained with Hoechst 33342. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Immunocytochemistry-Immunofluorescence-NBP2-42225-img0005.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - Human peripheral blood cells were stained (2 x 10^6 cells/mL) using the anti-CD63 antibody (Blue) at a dilution of 1:1000. Signal was detected using a Gt x Ms DyLight 488 Secondary and gated to the monocyte/granulocyte cell populations. Isotype was Mouse IgG1 kappa (orange). Data collected on BD FACS Calibur flow cytometer.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0003.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - A431 cells were stained (1 x10^6 cells/mL) using the anti-CD63 antibody at a 1:1000 dilution (blue). Signal was detected with Gt x Ms DyLight 488 secondary. Isotype control (orange).](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0004.jpg)
![Flow (Intracellular): CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on HepG2 cells with CD63 Antibody (H5C6) NBP2-42225AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Intracellular-NBP2-42225-img0009.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on HeLa cells with CD63 Antibody (H5C6) NBP2-42225APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Allophycocyanin.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0010.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on SK-MEL-28 cells with CD63 Antibody (H5C6) NBP2-42225AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0011.jpg)
![Flow Cytometry: CD63 Antibody (H5C6) [NBP2-42225] - An intracellular stain was performed on HeLa cells with CD63 [H5C6] Antibody NBP2-42225AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.](http://images.novusbio.com/fullsize/CD63-Antibody-H5C6-Flow-Cytometry-NBP2-42225-img0012.jpg)
![Western Blot: CD63 Antibody (H5C6) - BSA Free [NBP2-42225] - NCAM as a neuronal exosome marker in plasma. (A–C) Quantitative analysis of plasma EVs using flow cytometry. Dot plots of fluorescent intensity for plasma EVs stained with PE-CY7-labeled CD63 antibody. EVs were unstained (A) & stained with either isotype control (B) or CD63 antibody (C). (D) A18945 iPSC-derived neurons cultured for 6 weeks & immunostaining with mature neuronal markers (DAPI, MAP2, NeuN, & merged image). Images were taken using Zeiss LSM confocal microscope at ×63 magnification. Scale bar, 20 μm. (E) Representative images of cortical organoids at day 90 of differentiation. Scale bar, 1 mm. (F) The organoids express cortical layer marker BRN2 (also called POU3F2). The nuclei were stained with DAPI. Scale bar, 100 μm. (Gi) Markers for proliferating neural progenitors (SOX2) & cortical neuron marker CTIP2 (also known as BCL11B) with nuclei DAPI staining (Gii). (H) Western blot analysis of NCAM & CD63 from EVs released from iPSCs, iPSC-derived cortical neurons, & iPSC-derived brain organoids. EVs were isolated using SEC from the cell culture media of each sample, & equal EV particle numbers (6 × 108) were subject to immunoblotting with NCAM & CD63 antibodies. (I–L) Flow cytometry dot plots of fluorescent intensity for plasma EVs double-stained with DiI & NCAM antibody. EV samples were unstained (I), single stained with DiI (J), & double-stained with DiI & NCAM antibody in the absence (K) & presence (L) of 0.1% Triton X-100. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35655952), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-42225_mouse-monoclonal-cd63-antibody-h5c6-31020241612212.jpg)
![Western Blot: p73 Antibody (5B429) [NBP2-24737] - Consequences of DHX9 knockdown on protein levels of p53 targets in HCT116 p53+/+ and p53-/- cells. Western blot analysis of p53 transcriptional targets in HCT116 p53+/+ and p53-/- cells. Cells were harvested 6 days post-transduction and extracts were fractionated on 8% SDS-PAGE gels. All probings were performed on the same blot. Actin and eEF2 are used as loading controls. Quantitation of intensity levels of the proteins in the shDHX9 samples relative to the shFLuc. 1309 samples are indicated beneath each band. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/article/15889/text/), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Western-Blot-NBP2-24737-img0009.jpg "Western Blot: p73 Antibody (5B429) [NBP2-24737] - Consequences of DHX9 knockdown on protein levels of p53 targets in HCT116 p53+/+ and p53-/- cells. Western blot analysis of p53 transcriptional targets in HCT116 p53+/+ and p53-/- cells. Cells were harvested 6 days post-transduction and extracts were fractionated on 8% SDS-PAGE gels. All probings were performed on the same blot. Actin and eEF2 are used as loading controls. Quantitation of intensity levels of the proteins in the shDHX9 samples relative to the shFLuc. 1309 samples are indicated beneath each band. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/article/15889/text/), licensed under a CC-BY license.")
![Immunocytochemistry/Immunofluorescence: p73 Antibody (5B429) [NBP2-24737] - p73 was detected in immersion fixed Hela human cell line using p73 [5B429] Monoclonal Antibody (Catalog # NBP2-24737) at 3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Staining was observed in the cytoplasm and in the nuclei.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Immunocytochemistry-Immunofluorescence-NBP2-24737-img0008.jpg)
![Immunohistochemistry-Paraffin: p73 Antibody (5B429) [NBP2-24737] - FFPE breast cancer probed with p73 antibody at 5 ug/ml. Human tissue was used for this test. Staining of tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Immunohistochemistry-Paraffin-NBP2-24737-img0004.jpg)
![Flow Cytometry: p73 Antibody (5B429) [NBP2-24737] - Detection of p73 in Human HeLa Cell Line. Human HeLa cell line was stained with Mouse Anti- p73 Monoclonal Antibody (Catalog # NBP2-24737, filled histogram), or Mouse IgG1 isotype control (Catalog # MAB002, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Flow-Cytometry-NBP2-24737-img0007.jpg)
![Western Blot: p73 Antibody (5B429) [NBP2-24737] - Analysis of p73 in A) transfected cell lysate and B) HeLa cell lysate at 2 ug/ml.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Western-Blot-NBP2-24737-img0003.jpg)
![Flow (Intracellular): p73 Antibody (5B429) [NBP2-24737] - An intracellular stain was performed on HeLa cells with pan p73 (5B429) antibody NBP2-24737 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Flow-Intracellular-NBP2-24737-img0006.jpg)
![Simple Western: p73 Antibody (5B429) [NBP2-24737] - Lane view shows a specific band for p73 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.](http://images.novusbio.com/fullsize/p73-Antibody-5B429-Simple-Western-NBP2-24737-img0005.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - An intracellular stain was performed on HeLa cells with alpha-Smooth Muscle Actin Antibody (1A4/asm-1) NBP2-34522PCP (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to PerCP.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-17720241003238.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - An intracellular stain was performed on HeLa cells with alpha Smooth Muscle Actin (1A4/asm-1) Antibody NBP2-34522JF549 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Janelia Fluor 549.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-17720241044610.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - Human tonsil frozen tissue section stained for alpha-Smooth Muscle Actin (red) with the Janelia Fluor 549 conjugate of the Azide and BSA-Free version of alpha-Smooth Muscle Actin Antibody (1A4/asm-1), and counterstained with DAPI (blue). Image from verified customer review.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-17720249362624.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - An intracellular stain was performed on HeLa cells with alpha-Smooth Muscle Actin Antibody (1A4/asm-1) NBP2-34522PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-1772024936267.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - alpha-Smooth Muscle Actin expression in colorectal tumor cells. Using Allophycocyanin conjugated version of antibody (NBP2-34522APC). Image from verified customer review.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-1772024104461.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - An intracellular stain was performed on HeLa cells with alpha-Smooth Muscle Actin Antibody (1A4/asm-1) NBP2-34522AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-17720241003226.jpg)
![Flow Cytometry: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - Analysis of alpha-Smooth Muscle Actin Antibody (1A4/asm-1) [Alexa Fluor® 647] (Catalog Number: NBP2-34522AF647) using two lung cancer cell lines to acquire the subcutaneous tumors. MMT status of macrophages was then analyzed. Image from a verified customer review.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-110202413103111.png)
![Immunohistochemistry-Paraffin: alpha-Smooth Muscle Actin Antibody (1A4/asm-1) - BSA Free [NBP2-33006] - aSMA stained over night in human FFPE tonsil section ((Catalog # NBP2-33006AF750)). Heat mediated antigen retrieval at pH 9. 1:50 Dilution in 3% BSA, incubation over night at 4 °C. Image from a verified customer review.](http://images.novusbio.com/fullsize/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-61202511214025.png)
![Immunohistochemistry-Paraffin: Mouse Monoclonal alpha-Smooth Muscle Actin Antibody (1A4/asm-1) [NBP2-33006] - Images demonstrating SMA immunoreactivity in a variety of human FFPE tissue sections. NBP2-33006 was used at a concentration of 125ng/mL and was left on tissue sections for 30m at room temperature. Formalin fixed paraffin embedded sections were deparaffinized and rehydrated. Sections then underwent heat induced epitope retrieval in a citrate-based solution for 5min in a food steamer. Endogenous peroxidase activity was blocked by incubating slides for 15min in tris buffered saline containing 3% (volume/volume) hydrogen peroxide. Tissue sections were then washed, blocked with normal horse serum for 20 min, and then incubated with the primary antibody (125ng/mL) for 30min at room temperature. Sections were then washed and incubated with a horse anti-mouse HRP polymer for 30m at room temperature. Sections were washed and incubated with DAB chromogen for approximately 2.5min. Sections were then washed, counterstained with hematoxylin, dehydrated, cleared and mounted with a coverslip. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-immunohistochemistry-paraffin-2242025134951..png)
![Immunohistochemistry-Paraffin: Mouse Monoclonal alpha-Smooth Muscle Actin Antibody (1A4/asm-1) [NBP2-33006] - Images demonstrating SMA immunoreactivity in a variety of canine FFPE tissue sections. NBP2-33006 was used at a concentration of 125ng/mL and was left on tissue sections for 30m at room temperature. Formalin fixed paraffin embedded sections were deparaffinized and rehydrated. Sections then underwent heat induced epitope retrieval in a citrate-based solution for 5min in a food steamer. Endogenous peroxidase activity was blocked by incubating slides for 15min in tris buffered saline containing 3% (volume/volume) hydrogen peroxide. Tissue sections were then washed, blocked with normal horse serum for 20 min, and then incubated with the primary antibody (125ng/mL) for 30min at room temperature. Sections were then washed and incubated with a horse anti-mouse HRP polymer for 30m at room temperature. Sections were washed and incubated with DAB chromogen for approximately 2.5min. Sections were then washed, counterstained with hematoxylin, dehydrated, cleared and mounted with a coverslip. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-immunohistochemistry-paraffin-2242025135659..png)
![Immunohistochemistry-Paraffin: Mouse Monoclonal alpha-Smooth Muscle Actin Antibody (1A4/asm-1) [NBP2-33006] - Images demonstrating SMA immunoreactivity in a variety of feline FFPE tissue sections. NBP2-33006 was used at a concentration of 125ng/mL and was left on tissue sections for 30m at room temperature. Formalin fixed paraffin embedded sections were deparaffinized and rehydrated. Sections then underwent heat induced epitope retrieval in a citrate-based solution for 5min in a food steamer. Endogenous peroxidase activity was blocked by incubating slides for 15min in tris buffered saline containing 3% (volume/volume) hydrogen peroxide. Tissue sections were then washed, blocked with normal horse serum for 20 min, and then incubated with the primary antibody (125ng/mL) for 30min at room temperature. Sections were then washed and incubated with a horse anti-mouse HRP polymer for 30m at room temperature. Sections were washed and incubated with DAB chromogen for approximately 2.5min. Sections were then washed, counterstained with hematoxylin, dehydrated, cleared and mounted with a coverslip. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-immunohistochemistry-paraffin-224202514040..png)
![Immunohistochemistry-Paraffin: Mouse Monoclonal alpha-Smooth Muscle Actin Antibody (1A4/asm-1) [NBP2-33006] - SMA in FFPE sections of pig tissue. Formalin fixed paraffin embedded sections were deparaffinized and rehydrated. Sections then underwent heat induced epitope retrieval in a citrate-based solution for 5min in a food steamer. Endogenous peroxidase activity was blocked by incubating slides for 15min in tris buffered saline containing 3% (volume/volume) hydrogen peroxide. Tissue sections were then washed, blocked with normal horse serum for 20 min, and then incubated with the primary antibody (125ng/mL) for 30min at room temperature. Sections were then washed and incubated with a horse anti-mouse HRP polymer for 30m at room temperature. Sections were washed and incubated with DAB chromogen for approximately 2.5min. Sections were then washed, counterstained with hematoxylin, dehydrated, cleared and mounted with a coverslip. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp2-33006_mouse-monoclonal-alpha-smooth-muscle-actin-antibody-1a4-asm-1-immunohistochemistry-paraffin-224202514429..png)
![Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody - BSA Free [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - MEF were treated with 10 uM BA or DMSO (0.1%) for 16 h before they were subjected to a glycolysis stress test as described under](http://images.novusbio.com/fullsize/Pyruvate-Dehydrogenase-E1-alpha-subunit-[p-Ser293]-Antibody-Western-Blot-NB110-93479-img0007.jpg "<b>Biological Strategies Validation. </b>Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody - BSA Free [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - MEF were treated with 10 uM BA or DMSO (0.1%) for 16 h before they were subjected to a glycolysis stress test as described under")
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] antibody (1:250) was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).](http://images.novusbio.com/fullsize/Pyruvate-Dehydrogenase-E1-alpha-subunit-[p-Ser293]-Antibody-Immunocytochemistry-Immunofluorescence-NB110-93479-img0006.jpg)
![Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Detection of PDHE1 alpha [S293] in an in vitro autophosphorylation of PDH complex in response to different stimulants: (-) none, H202-hydrogen peroxide, AR-anhydroretinol, Rol-retinol, RA-retinoic acid, PMA, Lipid-PKC lipid activator. Photo courtesy of Dr. Beatrice Hoyos, Memorial Sloan-Kettering Cancer Center.](http://images.novusbio.com/fullsize/Pyruvate-Dehydrogenase-E1-alpha-subunit-[p-Ser293]-Antibody-Western-Blot-NB110-93479-img0003.jpg)
![Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - PDH activity and protein levels of PDK4 and p-PDH in hindlimb skeletal muscle. Samples were resolved by electrophoresis, transferred and probed by antibodies and representative immunoblots and normalization of pPDH are presented. The following comparisons were made: within a genotype, fed versus fasted; across genotypes, fed versus fed and fasted versus fasted. Bars that do not share a common symbol differ significantly (P < 0.05). Data presented as mean +/- SEM (n = 6). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124204), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Pyruvate-Dehydrogenase-E1-alpha-subunit-[p-Ser293]-Antibody-Western-Blot-NB110-93479-img0008.jpg)
![Western Blot: Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody - BSA Free [NB110-93479] - PDK4 expression during syncytialization of human placental trophoblasts. (A) Changes of the RPKM values for PDK family members in trophoblasts before (red column, 3 hours) & after (blue column, 48 hours) syncytialization. (B) Changes in PDK4 mRNA (black column, n = 4) & protein (white column, n = 4) abundance during syncytialization. (C) Changes in the phosphorylation of PDHE1 alpha during syncytialization n = 4. (D) Representative images showing intense staining of PDK4 (red) in the cytotrophoblast layer & weak staining of PDK4 in the syncytial layer of human chorionic villi at early gestation. The syncytial & cytotrophoblast layers were labeled with beta -hCG (green) & SPINT1 (green) respectively. Nuclei were counterstained with DAPI (blue). n = 3; *P < 0.05; **P < 0.01; ***P < 0.001 against 3 hours; n.s., not significant. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-09163-8), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb110-93479_rabbit-polyclonal-pyruvate-dehydrogenase-e1-alpha-subunit-p-ser293-antibody-310202416212313.jpg)
![Western Blot: EGLN3/PHD3 Antibody [NB100-139] - PHD3 is required for SCC cell survival in prolonged hypoxia. PHD3 is induced in hypoxia. Knock-down of PHD3 expression with siRNA (siPHD3) is specific and does not affect PHD2 or HIF-1 alpha expression. siScr = scrambled control siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0014617), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Knockdown-Validated-NB100-139-img0017.jpg "<b>Genetic Strategies Validation. </b>Western Blot: EGLN3/PHD3 Antibody [NB100-139] - PHD3 is required for SCC cell survival in prolonged hypoxia. PHD3 is induced in hypoxia. Knock-down of PHD3 expression with siRNA (siPHD3) is specific and does not affect PHD2 or HIF-1 alpha expression. siScr = scrambled control siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0014617), licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Immunohistochemistry-Paraffin: EGLN3/PHD3 Antibody [NB100-139] - Analysis of a formalin fixed tissue section of human renal cell carcinoma (clear cell type) using rabbit polyclonal EGLN3/PHD3 antibody with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a strong nuclear staining of PHD3 primarily in the cancer cells while the stromal cells were largely negative for this protein.](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Immunohistochemistry-Paraffin-NB100-139-img0010.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: EGLN3/PHD3 Antibody [NB100-139] - EGLN3/PHD3 depletion increases S10 phosphorylation of p27 in hypoxia. S10 phosphorylated form of p27 is strongly induced in EGLN3/PHD3 depleted cells. Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/s12943-015-0410-5) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Knockdown-Validated-NB100-139-img0019.jpg)
![Immunocytochemistry/Immunofluorescence: EGLN3/PHD3 Antibody [NB100-139] - EGLN3/PHD3 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Immunocytochemistry-Immunofluorescence-NB100-139-img0014.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: EGLN3/PHD3 Antibody [NB100-139] - BxPC3 cells stably transduced with retrovirus containing PHD3Wt (BxPC3-Wt), Vector (BxPC3-Vec) or anti-PHD3 shRNA (BxPC3-KD) were harvested for RNA and protein following 24 hours exposure to normoxia (21% O2) or hypoxia (1% O2). Whole cell lysate from BxPC3-Wt, BxPC3-Vec and BxPC3-KD cells following 24 hours exposure to normoxia (N) or hypoxia (H) was resolved by SDS-PAGE and blotted for actin (top) and PHD3 (bottom). PHD3 band intensity was quantified relative to actin in each lane and then normalized to BxPC3-Vec (N). Relative band intensity is indicated below the figure. NS = non-specific band running just above PHD3 band. Data is representative of >3 biological replicates. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0083021), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Knockdown-Validated-NB100-139-img0016.jpg)
![Western Blot: EGLN3/PHD3 Antibody [NB100-139] - The HIF-1 response is not impaired in prostate and breast carcinoma cell lines that lack PHD3. Melanoma, breast and prostate cancer cell lines with methylated PHD3 promoters (MB-435, PC-3) and non-methylated promoters (MCF7, DU 145) were subjected to hypoxia (1% O2) or normoxia for 24 hours. Thirty micrograms of whole cell lysate was western blotted for the presence of PHD3; actin was used as a loading control. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0014617), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Western-Blot-NB100-139-img0018.jpg)
![Western Blot: EGLN3/PHD3 Antibody [NB100-139] - Analysis of HeLa lysates using NB100-139. Image courtesy of Dr Gregg Semenza (The Johns Hopkins University School of Medicine, Baltimore, MD USA)](http://images.novusbio.com/fullsize/EGLN3-PHD3-Antibody-Western-Blot-NB100-139-img0013.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - Cell cycle block in PHD3 depleted cells is p27-dependent. a Reduced cell proliferation as judged by reduced cell amount of hypoxic siPHD3-depleted HeLa cells is rescued with concomitant p27 knockdown. b Quantification of cell survival from four independent experiments with 4–5 optical views per experiment. Asterisk indicates significant difference (p = 0,05; n = 4). Protein expression was monitored with western blotting from parallel samples. c Knockdown of p27 restores the effect of PHD3 depletion on cell cycle. HeLa cells exposed to the indicated double knockdown followed by 6 h hypoxia & FACS analysis. d Quantification of cell cycle phases from three independent experiments using indicated double knockdown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-31020241535673.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - 5-dAzaC effect on PHD3 transcript (A) & protein (B) levels in HCT116 & DLD-1 CRC cells. HCT116 & DLD-1 cells were cultured in DMEM for 6, 24 & 48 h either in the absence or in the presence of 5-dAzaC at a concentration of 1.00 or 5.00 μM under hypoxic or normoxic conditions. After incubation the cells were used for total RNA isolation & protein isolation. Total RNA was reverse-transcribed, & PHD3 cDNA levels were determined by RQ-PCR relative quantification analysis. RQ-PCR results were standardized by the geometric mean of PBGD & hMRPL19 cDNA levels. PHD3 cDNA levels are expressed as a multiplicity of the respective controls. Each sample was determined in triplicate & results are presented as the mean ± SE from three experiments *P < 0.05. The cell protein was separated by 10% SDS-PAGE, & transferred to a membrane that was then immunoblotted with Rp anti - PHD3 Ab & incubated with goat anti-rabbit HRP-conjugated Ab. The membrane was then stripped & reblotted with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The band densitometry readings were normalized to GAPDH loading control. The ratio PHD3 to GAPDH for control was assumed to be 1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415525254.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - DNA methylation & expression level of the PHD3 gene in HCT116 & DLD-1 CRC cells. A. HCT116 & DLD-1 cells were cultured under normoxic or hypoxic (1% O2) conditions for 48 hrs. Cells were then used for DNA isolation followed by bisulfite modification. Methylation percentage of three DNA fragments within the PHD3 CpG island (Additional file 1, Additional file 2) in HCT116 & DLD-1 cells under hypoxic & normoxic conditions was determined by Real Time PCR amplification of bisulfite treated standard & cell line DNA, followed by comparison of their HRM profiles. B. Cells were cultured in DMEM either in hypoxic (1%O2) or normoxic conditions for 48 hrs. After incubation, the cells were used for total RNA isolation & reverse transcription. The PHD3 cDNA levels were determined by RQ-PCR relative quantification analysis. RQ-PCR results were standardized by the geometric mean of PBGD & hMRPL19 cDNA levels. PHD3 cDNA levels are expressed as a multiplicity of these cDNA copies in the cell line’s calibrator. C. Cells were cultured in DMEM either in hypoxic (1%O2) (H) or normoxic (N) conditions for 48 hrs. Cells were then used for protein isolation. Proteins were separated by 10% SDS-PAGE, & transferred to a membrane that was then immunoblotted with Rp anti - PHD3 Ab & incubated with goat anti-rabbit HRP-conjugated Ab. The membrane was then stripped & reblotted with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The band densitometry readings were normalized to GAPDH loading control. The ratio of PHD3 to GAPDH for DLD-1 in normoxic conditions was assumed to be 1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415532022.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - PHD3 depletion increases S10 phosphorylation of p27 in hypoxia. a Western blot analysis of p27 expression in cells arrested at different cell cycle phases & exposed to PHD3 or control siRNA. PHD3 depletion has no effect on T187 phosphorylation of p27 at any cell cycle phase. b PHD3 depletion has little effect on p27 phosphorylated on T157 & T198. c S10 phosphorylated form of p27 is strongly induced in PHD3 depleted cells. d Quantification of total & indicated p27 phosphoproteins from four independent experiments normalized to beta -actin. Out of the phosphoproteins the effect on the expression of p27S10 is most prominent. Asterisk indicates significant difference (p < 0,005; n = 4) (e) Quantification of indicated p27 phosphoproteins from four independent experiments normalized to total p27 expression. The effect on the expression of p27S10 is most prominent & comparable to the total p27 expression. Asterisk indicates significant difference (p < 0,05; n = 4). Only minor effect on T157 & T198 are seen Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-31020241682140.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - p27S10 phosphorylation is required for PHD3 depletion induced increase in p27 half-life. a Quantification of p27S10 phosphorylation from three independent experiments in 786-O cells. S10 phosphorylation demonstrated 2-fold increase at 6 h after cell cycle release in PHD3 depleted cells. b Serum starved 786-O cells were released from G0 block & S10 phosphorylation was monitored at the indicated timepoints under PHD3 depletion. PHD3 depletion had a prominent effect on S10 phosphorylation of p27. c Verification of subcellular localization of p27 plasmids. Immunofluorescence staining of Flag-p27wt / Flag-p27S10A transfected HeLa cells after 24 h hypoxia. As expected, p27wt is localized more into cytoplasmic than nuclear compartment. p27S10A is more localized into nucleus than into cytoplasm. Quantification of the subcellular localization from three optical fields (40x) (N; nuclear, C; cytoplasmic). d Flag-p27wt & S10A phosphorylation-deficient mutant (p27S10A) were transfected into siRNA-exposed HeLa cells. After 24 h of hypoxia CHX chase was performed. p27wt demonstrates strongly increased stability in siPHD3 exposed cells whereas p27S10A did not show any increased stability Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202416573.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - Experimental setup & the effect of PHD3 silencing on 786-O proteome. a siRNA-mediated silencing of PHD3 protein level in 786-O ccRCC cell line using two individual siRNA sequences in normoxic & in hypoxic (1% O2) condition. Quantification of three biological replicates, mean ± SEM, fold change to control (Scr). Asterisk indicates a statistically significant difference (*p < 0.05, **p < 0.01). b siRNA-mediated silencing of PHD3 mRNA expression, quantification of three individual experiments. Mean ± SEM, fold change to Scr (***p < 0.001). c Flow chart of the experimental procedure. 786-O cells were transfected with siPHD3#1 or with a non-targeting control siRNA (Scr) for 24 h followed by a hypoxic (1% O2) or normoxic (21% O2) exposure. Three independent experiments were performed; proteins were extracted, followed by in-gel digestion with trypsin. Purified peptides were ran through mass spectrometer. Protein identification was done with Mascot database search. Protein quantification was carried out with Progenesis QI, followed by testing the differential expression between sample groups using peptide-level expression change averaging (PECA). d Western blot validations of selected proteins in 786-O & RCC4 cell lines with two individual siRNA sequences targeting PHD3, representative analyses are shown Image collected & cropped by CiteAb from the following publication (//cancerandmetabolism.biomedcentral.com/articles/10.1186/s40170-017-0167-y), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-31020241682128.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 & subsequent release shows an increase of p27 expression in siPHD3 exposed cells. b Quantification for p27 expression under PHD3 depletion at indicated time points after cell cycle release in HeLa & 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 & inhibition of protein synthesis with cycloheximide indicate increased p27 stability in PHD3 depleted HeLa cells. d Quantification of p27 expression using siPHD3 or control at indicated time points. Four independent experiments (± SEM) are shown (p < 0,05; n = 4). e Analysis of p27 stability in 786–0 cells by cycloheximide chase during reoxygenation after 24 h hypoxia demonstrates markedly increased half-life of p27 upon PHD3 depletion Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415541995.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - PHD3 inhibition reduces the amount of hyperphosphorylated Rb & increases p27 in hypoxia.(A) SCC2 cells were transfected with the indicated siRNAs & exposed to normoxia or hypoxia for 24 to 48 hours followed by western blot analysis of PHD3, phosphorylated Rb (p-Rb) & cyclin B1. (B) HeLa cells were transfected with the indicated siRNAs, synchronized & exposed to normoxia or hypoxia for 6 to 24 hours after release. PHD3, phosphorylated Rb (p-Rb) & cyclin D1 were analyzed from samples by western blotting. (C) SCC2 cells were transfected with the indicated siRNAs & exposed to normoxia or hypoxia for 24 hours followed by western blot analysis of p21(Cip1). (D) Cells transfected with the indicated siRNAs & exposed to normoxia or hypoxia for 24 hours followed by western blot analysis of p16 & p27. Hypoxia was monitored by HIF-1a expression. (E) Cells transfected with the indicated siRNAs & exposed to normoxia or hypoxia for 24 & 48 hours followed by western blot analysis of p27. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22087251), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415532079.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - Cell cycle block under PHD3 depletion is accompanied by p27 induction. a PHD3 depletion induces a cell cycle block in G0/G1. HeLa & renal cell adenocarcinoma cells (786-O) were transfected with control (siScr) or PHD3 targeted (siPHD3) siRNA followed by synchronization at G0 & 24-h hypoxic exposure. Cell cycle progression was monitored by FACS analysis 8 h after cell cycle release. The combined means of three independent experiments are presented (±SEM) shown in the tables below. b PHD3 depletion induces p27 expression in HeLa cells & in 786-O cells under hypoxia (1 % O2) & normoxia (21 % O2) by siPHD3 & independent adenoviral shRNA against PHD3. p21 or p16 expression is not elevated by PHD3 knockdown. c Depletion of either PHD1 or PHD2 by siRNA does not increase p27 expression in 786-O cells Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415541957.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - p27S10 phosphorylation is required for PHD3 depletion induced increase in p27 half-life. a Quantification of p27S10 phosphorylation from three independent experiments in 786-O cells. S10 phosphorylation demonstrated 2-fold increase at 6 h after cell cycle release in PHD3 depleted cells. b Serum starved 786-O cells were released from G0 block & S10 phosphorylation was monitored at the indicated timepoints under PHD3 depletion. PHD3 depletion had a prominent effect on S10 phosphorylation of p27. c Verification of subcellular localization of p27 plasmids. Immunofluorescence staining of Flag-p27wt / Flag-p27S10A transfected HeLa cells after 24 h hypoxia. As expected, p27wt is localized more into cytoplasmic than nuclear compartment. p27S10A is more localized into nucleus than into cytoplasm. Quantification of the subcellular localization from three optical fields (40x) (N; nuclear, C; cytoplasmic). d Flag-p27wt & S10A phosphorylation-deficient mutant (p27S10A) were transfected into siRNA-exposed HeLa cells. After 24 h of hypoxia CHX chase was performed. p27wt demonstrates strongly increased stability in siPHD3 exposed cells whereas p27S10A did not show any increased stability Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415541913.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 & subsequent release shows an increase of p27 expression in siPHD3 exposed cells. b Quantification for p27 expression under PHD3 depletion at indicated time points after cell cycle release in HeLa & 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 & inhibition of protein synthesis with cycloheximide indicate increased p27 stability in PHD3 depleted HeLa cells. d Quantification of p27 expression using siPHD3 or control at indicated time points. Four independent experiments (± SEM) are shown (p < 0,05; n = 4). e Analysis of p27 stability in 786–0 cells by cycloheximide chase during reoxygenation after 24 h hypoxia demonstrates markedly increased half-life of p27 upon PHD3 depletion Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415533848.jpg)
![Western Blot: EGLN3/PHD3 Antibody - BSA Free [NB100-139] - Hypoxia induced dedifferentiation employs both HIF-dependent & independent mechanismsA. Mammosphere formation efficiency in MCF-7 cells transfected with siHIF1 alpha and/or siHIF2 alpha , & cultured in normoxia or hypoxia for 3 days. B. Percentage of ALDH+ cells in T47D cells transfected with siHIF1 alpha and/or siHIF2 alpha , & cultured in normoxia or hypoxia. C. Percentage of CD44+CD24−/low cells from MDA-MB-468 cells transfected with siHIF1 alpha and/or siHIF2 alpha , & cultured in normoxia or hypoxia. Data are presented as mean ±SEM of 8 independent experiments. (D, E) HIF1 alpha & HIF2 alpha mRNA D. & protein E. expression in CSCs & non-CSCs from MDA-MB-468 cells cultured in normoxia or hypoxia. F. Percentage of CD44+CD24−/low MDA-MB-468 cells grown in normoxia after silencing all three PHDs individually or collectively or in hypoxia. G. Protein expression of HIF1 alpha & HIF2 alpha in MDA-MB-468 cells transfected with a control siRNA or a siRNA directed to PHD3 & cultured in normoxia or hypoxia. H., I. PHD1, PHD2 & PHD3 mRNA H. & protein I. expression levels in CSCs & non-CSCs sorted from MDA-MB-468 cells. B, D, F & H show means ±SD of three independent experiments. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.5564), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-139_rabbit-polyclonal-egln3-phd3-antibody-310202415525283.jpg)
![Immunocytochemistry/Immunofluorescence: Nanog Antibody [NB100-58842] - The stem cell marker (Nanog) staining of iPS-BP cells. Nuclei were stained with DAPI (blue). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0044024) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunocytochemistry-Immunofluorescence-NB100-58842-img0015.jpg "Immunocytochemistry/Immunofluorescence: Nanog Antibody [NB100-58842] - The stem cell marker (Nanog) staining of iPS-BP cells. Nuclei were stained with DAPI (blue). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0044024) licensed under a CC-BY license.")
![Immunohistochemistry-Paraffin: Nanog Antibody [NB100-58842] - Nanog-inducible mouse model. Immunohistochemistry (IHC) for NANOG of paraffin-embedded sections of tail skin from CTR and TG mice treated as indicated in (B). Two magnifications are shown for each tissue (bars correspond to 50 um). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep10205), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunohistochemistry-Paraffin-NB100-58842-img0011.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - Whole cell lysate (5, 15, and 50 ug) from F9 cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Nanog antibody used for WB at 0.5 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.](http://images.novusbio.com/fullsize/Nanog-Antibody-Western-Blot-NB100-58842-img0009.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody [NB100-58842] - Nanog-inducible mouse model. Representative hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) for NANOG, Ki67 and LORICRIN of TPA-treated CTR and TG mice (bars correspond to 50 um). Statistical significance was determined by the two-tailed Student's t test: (*) p < 0.05; (**) p < 0.01. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep10205), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunohistochemistry-Paraffin-NB100-58842-img0012.jpg)
![Immunocytochemistry/Immunofluorescence: Nanog Antibody [NB100-58842] - Nanog staining in ES cells treatment with leukemia inhibitory factor (LIF). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0044024) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunocytochemistry-Immunofluorescence-NB100-58842-img0013.jpg)
![Immunohistochemistry: Nanog Antibody [NB100-58842] - Differentiation of wt and Suv4-20dn teratomas. Representative images of Nanog staining in wt and Suv4-20dn teratomas. Scale bar, 200 um. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0025680) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunohistochemistry-NB100-58842-img0016.jpg)
![Immunoprecipitation: Nanog Antibody [NB100-58842] - Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from F9 cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Nanog antibody used for IP at 6 ug per reaction. Nanog was also immunoprecipitated by rabbit anti-Nanog antibody. For blotting immunoprecipitated Nanog, this was used at 1 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunoprecipitation-NB100-58842-img0010.jpg)
![<b>Genetic Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Nanog Antibody [NB100-58842] - Nanog antibody was tested in DGCR8 knockout Mouse embryonic stem cells (NBA1-19349) with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunocytochemistry-Immunofluorescence-NB100-58842-img0004.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - The forced expression of Tlk1 results in the aberrant downregulation of core pluripotency factors & attenuates self-renewal. (A) Immunoblot analysis of Oct4, Sox2, & Nanog levels in control mESCs (empty vector or doxycycline depletion) & Tlk1-overexpressing mESCs. The mESCs expressing an empty vector or the Tet-On-Tlk1 or Tet-On-Tlk1-D607A expression vector were cultured in the absence or presence of doxycycline (Dox; 100 ng/ml) for 24 hrs under undifferentiated self-renewal conditions. (B) Quantification of results from (A). The protein levels of the target genes were normalized to alpha -tubulin levels. The protein expression levels of each mESC line not treated with doxycycline were normalized to 1. The biological data are presented as mean (n = 6) ± SEM. *Р < 0.05, & **P < 0.01. (C) The morphology & AP staining of Tet-On-inducible Tlk1-expressing cell lines cultured in mock (Dox−) or doxycycline (Dox+) for 48 hrs. Scale bar, 500 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-31020241554197.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - Tlk1-deficiency in mESCs causes a delay in the downregulation of core pluripotency factors upon differentiation. (A,C & E) Representative immunoblotting images showing Tlk1, Oct4, Sox2, & Nanog levels in Tlk1-KD cells upon differentiation. Differentiation was induced three different ways as previously described in Fig. 3. Alpha-tubulin was used as the loading control. (B,D & F) Quantification of the relative expression of the target proteins in panels (A,C, & E). The target proteins levels were normalized to that of alpha -tubulin. The protein expression levels of shLuc KD cells were normalized to 1. The biological data are presented as mean (n = 4) ± SEM for LIF- & EB & for RA (n = 3). *Р < 0.05, **P < 0.01, & ***P < 0.001. (G) Immunofluorescence analysis of Oct4, Nanog & Tlk1 in control (shLuc) & Tlk1-deficient mESCs. Scale bars represent 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-310202415525256.jpg)
![Immunocytochemistry/ Immunofluorescence: Nanog Antibody [NB100-58842] - Tlk1-deficiency in mESCs causes a delay in the downregulation of core pluripotency factors upon differentiation. (A,C & E) Representative immunoblotting images showing Tlk1, Oct4, Sox2, & Nanog levels in Tlk1-KD cells upon differentiation. Differentiation was induced three different ways as previously described in Fig. 3. Alpha-tubulin was used as the loading control. (B,D & F) Quantification of the relative expression of the target proteins in panels (A,C, & E). The target proteins levels were normalized to that of alpha -tubulin. The protein expression levels of shLuc KD cells were normalized to 1. The biological data are presented as mean (n = 4) ± SEM for LIF- & EB & for RA (n = 3). *Р < 0.05, **P < 0.01, & ***P < 0.001. (G) Immunofluorescence analysis of Oct4, Nanog & Tlk1 in control (shLuc) & Tlk1-deficient mESCs. Scale bars represent 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-310202415525252.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - Tlk1 is not required for mESC self-renewal or pluripotency. (A) The efficiency of Tlk1 knockdown (KD) in control (shLuc) & Tlk1-KD mESCs (shTlk1 #1 & #2) was confirmed by RT-qPCR analysis. Data are mean (n = 3) ± SEM. **Р < 0.01 & ***Р < 0.001. (B) The morphology of control (shLuc) & Tlk1-KD (shTlk1 #1 & #2) mESCs was evaluated using phase-contrast microscopic images & AP staining. Scale bars represent 500 µm. (C & D) The mRNA expression of pluripotency-associated & development-associated genes were analyzed by RT-qPCR in control (shLuc) & Tlk1-KD (shTlk1 #1 & #2) mESCs cultured under undifferentiated self-renewal conditions. All data were normalized to Gapdh & plotted relative to the expression level in control cells. Data are means (n = 3) ± SEM. *Р < 0.05, **Р < 0.01, & ***Р < 0.001. (E) The protein levels of pluripotency factors in control (shLuc) & Tlk1-KD (shTlk1 #1 & #2) mESCs was analyzed by immunoblotting using antibodies specific to Oct4, Sox2, & Nanog. (F) Quantification based on densitometry of Western blotting data from (E). All data were normalized to alpha -tubulin. Data are means (n = 3) ± SEM. *Р < 0.05, **Р < 0.01, & ***Р < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-310202415532014.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - Tlk1-deficiency in mESCs causes a delay in the downregulation of core pluripotency factors upon differentiation. (A,C & E) Representative immunoblotting images showing Tlk1, Oct4, Sox2, & Nanog levels in Tlk1-KD cells upon differentiation. Differentiation was induced three different ways as previously described in Fig. 3. Alpha-tubulin was used as the loading control. (B,D & F) Quantification of the relative expression of the target proteins in panels (A,C, & E). The target proteins levels were normalized to that of alpha -tubulin. The protein expression levels of shLuc KD cells were normalized to 1. The biological data are presented as mean (n = 4) ± SEM for LIF- & EB & for RA (n = 3). *Р < 0.05, **P < 0.01, & ***P < 0.001. (G) Immunofluorescence analysis of Oct4, Nanog & Tlk1 in control (shLuc) & Tlk1-deficient mESCs. Scale bars represent 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-310202415532036.jpg)
![Western Blot: Nanog Antibody [NB100-58842] - Tlk1-deficiency in mESCs causes a delay in the downregulation of core pluripotency factors upon differentiation. (A,C & E) Representative immunoblotting images showing Tlk1, Oct4, Sox2, & Nanog levels in Tlk1-KD cells upon differentiation. Differentiation was induced three different ways as previously described in Fig. 3. Alpha-tubulin was used as the loading control. (B,D & F) Quantification of the relative expression of the target proteins in panels (A,C, & E). The target proteins levels were normalized to that of alpha -tubulin. The protein expression levels of shLuc KD cells were normalized to 1. The biological data are presented as mean (n = 4) ± SEM for LIF- & EB & for RA (n = 3). *Р < 0.05, **P < 0.01, & ***P < 0.001. (G) Immunofluorescence analysis of Oct4, Nanog & Tlk1 in control (shLuc) & Tlk1-deficient mESCs. Scale bars represent 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29321513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-58842_rabbit-polyclonal-nanog-antibody-31020241683922.jpg)
![Western Blot: YAP1 Antibody (2F12) - Isoform 5 [H00010413-M01] - YAP1 Antibody (2F12) [H00010413-M01] - JNK phosphorylation of YAP enhances the stabilization of delta-Np63alpha by direct binding. H357 cells were transfected with control siRNA (siCTL) or YAP siRNA (siYAP) and 72 h later the cells were harvested for IP of endogenous p63. p63 IP eluates and input lysates were analyzed by Western blotting using the indicated antibodies. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis20107) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Knockdown-Validated-H00010413-M01-img0016.jpg "<b>Genetic Strategies Validation. </b>Western Blot: YAP1 Antibody (2F12) - Isoform 5 [H00010413-M01] - YAP1 Antibody (2F12) [H00010413-M01] - JNK phosphorylation of YAP enhances the stabilization of delta-Np63alpha by direct binding. H357 cells were transfected with control siRNA (siCTL) or YAP siRNA (siYAP) and 72 h later the cells were harvested for IP of endogenous p63. p63 IP eluates and input lysates were analyzed by Western blotting using the indicated antibodies. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis20107) licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Western Blot: YAP1 Antibody (2F12) [H00010413-M01] - Analysis of YAP1 Antibody (2F12) Isoform 5 expression in Hela S3 N cells. Observed molecular weight 75 kDa.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Western-Blot-H00010413-M01-img0012.jpg)
![Immunocytochemistry/Immunofluorescence: YAP1 Antibody (2F12) [H00010413-M01] - Analysis of monoclonal antibody to YAP1 on HeLa cell. Antibody concentration 30 ug/ml.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Immunocytochemistry-Immunofluorescence-H00010413-M01-img0009.jpg)
![Immunohistochemistry-Paraffin: YAP1 Antibody (2F12) [H00010413-M01] - Analysis of monoclonal antibody to YAP1 on formalin-fixed paraffin-embedded human endometrium. Antibody concentration 3 ug/ml.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Immunohistochemistry-Paraffin-H00010413-M01-img0011.jpg)
![Western Blot: YAP1 Antibody (2F12) [H00010413-M01] - JNK specifically phosphorylates YAP in vivo. HaCaT, U2OS, MCF7 and 293T cell lines were treated with anisomycin or DMSO (control) for 60 min before harvesting. YAP band shift was analyzed as described before and lysates were analyzed by Western blotting with the indicated antibodies. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis20107) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Western-Blot-H00010413-M01-img0017.jpg)
![Western Blot: YAP1 Antibody (2F12) [H00010413-M01] - Immunogen detection using YAP1 antibody (2F12) Isoform 5, observered MW 37.73KDa) .](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Western-Blot-H00010413-M01-img0019.jpg)
![Western Blot: YAP1 Antibody (2F12) [H00010413-M01] - Analysis of YAP1 Antibody (2F12) Isoform 5 in transfected 293T cell line. Lane 1: YAP1 transfected lysate (TMW: 54.5 KDa, observed MW 75 KDa). Lane 2: Non-transfected lysate.](http://images.novusbio.com/fullsize/YAP1-Antibody-2F12-Western-Blot-H00010413-M01-img0018.jpg)