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 | NB110-94158SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-25164SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP1-48308SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP1-87679-25ul | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP1-25959SS | |||
| Azide and BSA Free | NBP2-80902 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF4709-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF1865-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | IC317G-100UG | |||
| Carrier Free | IC317P | |||
| Carrier Free | IC317A | |||
| Carrier Free | MAB317-SP | |||
| Carrier Free | MAB317-500 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB200-111SS | |||
![Western Blot: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - CLDN1 monoclonal antibody (M01), clone 1C5-D9. Analysis of CLDN1 expression in A-431.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Western-Blot-H00009076-M01-img0015.jpg "Western Blot: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - CLDN1 monoclonal antibody (M01), clone 1C5-D9. Analysis of CLDN1 expression in A-431.")
![Immunohistochemistry-Paraffin: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - Analysis of monoclonal antibody to CLDN1 on formalin-fixed paraffin-embedded human colon. Antibody concentration 5 ug/ml.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Immunohistochemistry-Paraffin-H00009076-M01-img0011.jpg)
![Western Blot: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - CLDN1 monoclonal antibody (M01), clone 1C5-D9. Analysis of CLDN1 expression in PRC/PRF/5.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Western-Blot-H00009076-M01-img0014.jpg)
![Western Blot: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - Analysis of CLDN1 expression in transfected 293T cell line by CLDN1 monoclonal antibody (M01), clone 1C5-D9.Lane 1: CLDN1 transfected lysate(23 KDa).Lane 2: Non-transfected lysate.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Western-Blot-H00009076-M01-img0016.jpg)
![Immunohistochemistry-Paraffin: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - Analysis of monoclonal antibody to CLDN1 on formalin-fixed paraffin-embedded human small Intestine. Antibody concentration 3 ug/ml.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Immunohistochemistry-Paraffin-H00009076-M01-img0009.jpg)
![Immunoprecipitation: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - Analysis of CLDN1 transfected lysate using anti-CLDN1 monoclonal antibody and Protein A Magnetic Bead, and immunoblotted with CLDN1 MaxPab rabbit polyclonal antibody.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-Immunoprecipitation-H00009076-M01-img0012.jpg)
![ELISA: Claudin-1 Antibody (1C5-D9) [H00009076-M01] - Detection limit for recombinant GST tagged CLDN1 is approximately 0.03ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/Claudin-1-Antibody-1C5-D9-ELISA-H00009076-M01-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: Mre11 Antibody (12D7) [NB100-473] - Human primary cells stained with Mre11 antibody at a dilution of 1:60. cells were fixed in 1:1 Acetone/Methanol. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Immunohistochemistry-NB100-473-img0008.jpg "Immunocytochemistry/Immunofluorescence: Mre11 Antibody (12D7) [NB100-473] - Human primary cells stained with Mre11 antibody at a dilution of 1:60. cells were fixed in 1:1 Acetone/Methanol. ICC/IF image submitted by a verified customer review.")
![<b>Genetic Strategies Validation. </b>Western Blot: Mre11 Antibody (12D7) [NB100-473] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Mre11 antibody [12D7] diluted at 1:500. HRP-conjugated anti-mouse IgG antibody (NBP2-19382) was used to detect the primary antibody. A band appears at the theoretical molecular weight of 80 kDa.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Western-Blot-NB100-473-img0010.jpg)
![Western Blot: Mre11 Antibody (12D7) [NB100-473] - Mre11 antibody [12D7] detects Mre11 protein by western blot analysis. A. 30 ug 293T whole cell extract. B. 30 ug whole cell extract of human Mre11-transfected 293T cells. 7.5% SDS-PAGE. Mre11 antibody [12D7] (NB100-473). HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody. Bands are prominent at the theoretical molecular weight of 80 kDa.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Western-Blot-NB100-473-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: Mre11 Antibody (12D7) [NB100-473] - Analysis of Mre11 antibody (12D7) on lymphoblastoid cells. Dilution 1:200. Image from verified customer review.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Immunocytochemistry-Immunofluorescence-NB100-473-img0021.jpg)
![Immunocytochemistry/Immunofluorescence: Mre11 Antibody (12D7) [NB100-473] - Mre11 antibody [12D7] detects Mre11 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: Mre11 stained by Mre11 antibody [12D7] (NB100-473).Blue: Hoechst 33342 staining.Scale bar= 10 um.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Immunocytochemistry-Immunofluorescence-NB100-473-img0015.jpg)
![Immunocytochemistry/Immunofluorescence: Mre11 Antibody (12D7) [NB100-473] - Mouse fibroblasts stained with Mre11 Antibody (12D7). ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/Mre11-Antibody-12D7-Immunocytochemistry-Immunofluorescence-NB100-473-img0020.jpg)
![Western Blot: Mre11 Antibody (12D7) [NB100-473] - Various whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Mre11 antibody [12D7] (NB100-473) diluted at 1:500. The HRP-conjugated anti-mouset IgG antibody was used to detect the primary antibody.](http://images.novusbio.com/fullsize/nb100-473_mouse-monoclonal-mre11-antibody-12d7-91020242091437.jpg)
![Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - HepG2 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-MAT2A Antibody (NB110-94158) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 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/MAT2A-Antibody-Immunocytochemistry-Immunofluorescence-NB110-94158-img0010.jpg "Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - HepG2 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-MAT2A Antibody (NB110-94158) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.")
![Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - NIH3T3 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-MAT2A Antibody (NB110-94158) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 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/MAT2A-Antibody-Immunocytochemistry-Immunofluorescence-NB110-94158-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - PC12 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-MAT2A Antibody (NB110-94158) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 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/MAT2A-Antibody-Immunocytochemistry-Immunofluorescence-NB110-94158-img0012.jpg)
![Western Blot: MAT2A Antibody [NB110-94158] - Loss of PKM2 impairs methylation capacity, reduces DNA methylation and leads to the expression of endogenous retroviral elements. Western blot analysis of MAT2A, P53, P21 and PKM2 in control, P53KD, PKM2KD and PKM2KD/P53KD ECs. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-06406-8), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Western-Blot-NB110-94158-img0008.jpg)
![Immunohistochemistry-Paraffin: MAT2A Antibody [NB110-94158] - Analysis of a FFPE tissue section of human kidney using 1:200 dilution of MAT2A antibody (NB110-94158). The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Immunohistochemistry-Paraffin-NB110-94158-img0009.jpg)
![Western Blot: MAT2A Antibody [NB110-94158] - Total protein from HeLa and HepG2 cells and Human and Mouse Kidney was separated on a 12% 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-MAT2A in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Western-Blot-NB110-94158-img0005.jpg)
![Western Blot: MAT2A Antibody [NB110-94158] - Detection of MAT2A in HepG2 whole cell lysates.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Western-Blot-NB110-94158-img0003.jpg)
![Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - MAT2A antibody was tested in HepG2 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).](http://images.novusbio.com/fullsize/MAT2A-Antibody-Immunocytochemistry-Immunofluorescence-NB110-94158-img0004.jpg)
![Immunocytochemistry/Immunofluorescence: MAT2A Antibody [NB110-94158] - Mouse embryonic stem cells (mESCs) were fixed with paraformaldehyde, permeabilized with Triton, blocked with 4% BSA and 1% normal goat serum, and incubated with 1:200 dilution of MAT2A antibody. Then, samples were stained with anti-rabbit Alexa Fluor 594 (1:1000) and NucBlue. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Immunocytochemistry-Immunofluorescence-NB110-94158-img0006.jpg)
![Simple Western: MAT2A Antibody [NB110-94158] - Human HepG2 cell lysate (0.2 ug/uL total protein). Antibody at 1:250. Simple Western image submitted by a verified customer review.](http://images.novusbio.com/fullsize/MAT2A-Antibody-Simple-Western-NB110-94158-img0007.jpg)
![Immunocytochemistry/ Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Validation of UBQLN2 antibody for staining of endogenous UBQLN2 in U2OS cells. U2OS cells were transfected with control siRNA or siRNA targeting UBQLN2. Cells were either seeded as separate groups (i.e., control and UBQLN2 siRNA) or mixed 1:1 and seeded together. Cells were stained using the mouse monoclonal anti-UBQLN2 6H9 (Novus NBP2-25164), at 1:250 in 2% BSA PBS for 1h. Secondary antibody was goat Anti-Mouse DyLight 488. Knockdown of UBQLN2 can be clearly seen to decrease the signal, indicating that the antibody is specific to UBQLN2. (G) displays zoom of the indicated areas in. For the mixed cells a white arrowhead indicates a cell transfected with control siRNA and a black arrowhead a cell transfected with UBQLN2 siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1016/j.cell.2016.07.001), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockdown-Validated-NBP2-25164-img0010.jpg "<b>Genetic Strategies Validation. </b>Immunocytochemistry/ Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Validation of UBQLN2 antibody for staining of endogenous UBQLN2 in U2OS cells. U2OS cells were transfected with control siRNA or siRNA targeting UBQLN2. Cells were either seeded as separate groups (i.e., control and UBQLN2 siRNA) or mixed 1:1 and seeded together. Cells were stained using the mouse monoclonal anti-UBQLN2 6H9 (Novus NBP2-25164), at 1:250 in 2% BSA PBS for 1h. Secondary antibody was goat Anti-Mouse DyLight 488. Knockdown of UBQLN2 can be clearly seen to decrease the signal, indicating that the antibody is specific to UBQLN2. (G) displays zoom of the indicated areas in. For the mixed cells a white arrowhead indicates a cell transfected with control siRNA and a black arrowhead a cell transfected with UBQLN2 siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1016/j.cell.2016.07.001), licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Simple Western: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Simple Western lane view shows a specific band for Ubiquilin 2 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Simple-Western-NBP2-25164-img0007.jpg)
![Western Blot: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of different tissue and cell lysates using mouse mAb to ubiquilin 2, NBP2-25164, dilution 1:1,000 in green: [1] protein standard (red), [2] NIH-3T3, [3] C6, [4] HEK293, [5] HeLa, [6] SH-SY5Y, [7] rat whole brain, and [8] mouse whole brain. The band at 65-70kDa corresponds to ubiquilin 2 protein, which is known to differ between the human and rodent proteins.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Western-Blot-NBP2-25164-img0009.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of an NIH-3T3 cell culture stained with mouse mAb to ubiquilin 2, NBP2-25164, dilution 1:1,000 in green, and costained with chicken pAb to lamin A/C, dilution 1:5,000 in red. The blue is DAPI staining of nuclear DNA. The cells were treated with 50uM of chloroquine, an inhibitor of autophagy, for 16 hours prior to staining. NBP2-25164 antibody reveals vesicular staining of ubiquilin 2 protein accumulated in lysosomes in the cytoplasm, while the lamin A/C antibody stains the nuclear lamina.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0008.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of Human U2OS cells stained with Ubiquilin 2 antibody. Primary antibody dilution: 1:1000. Image from verified customer review.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HeLa cell cultures stained with NBP2-25164 (green) and chicken polyclonal antibody to Vimentin: NB300-223 (red). In most individual cells Ubiquilin 2 is present diffusely in the cytoplasm of cells, though some cells show enrichment of the protein in spherical autophagosome-like structure.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0003.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Lysates of HAP1 WT and Ubiquilin 2 KO were prepared, and 30 ug of protein were processed for immunoblot with NBP2-25164. The Ponceau stained transfer of the blot is shown. Antibody dilution used: 1/2000. Predicted band size: 65 kDa. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockout-Validated-NBP2-25164-img0012.jpg)
![Immunoprecipitation: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HAP1 lysates were prepared, and immunoprecipitation was performed using 2.0 ug of Ubiquilin 2 antibody (NBP2-25164) pre-coupled to Dynabeads protein G. Samples were washed and processed for immunoblot with NBP2-25164. For immunoblot, Recombinant Ubiquilin 2 (Rb x Ubiquilin 2) was used at 1/1000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunoprecipitation-NBP2-25164-img0013.jpg)
![<b>Genetic Strategies Validation. </b>Immunocytochemistry/ Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HAP1 WT and Ubiquilin 2 KO cells were labeled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with glass bottom. Cells were stained with Ubiquilin-2 antibody (NBP2-25164) and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative image of the blue and red (grayscale) channels is shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Antibody dilution used: 1/1000. Bars = 10 um Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockout-Validated-NBP2-25164-img0014.jpg)
![Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Expression profile of PKM isoforms in tissues from mouse organs. PKM1 and PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S2b. Results are presented as the mean +/- SD (** P < 0.01). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep08647) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0018.jpg "Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Expression profile of PKM isoforms in tissues from mouse organs. PKM1 and PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S2b. Results are presented as the mean +/- SD (** P < 0.01). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep08647) licensed under a CC-BY license.")
![Immunocytochemistry/Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in frozen sections of rat glioblastoma tissue. Image courtsey of anonymous customer review.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NBP1-48308-img0007.jpg)
![Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308] - Staining of PKM2 in mouse liver tissue.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Immunohistochemistry-NBP1-48308-img0009.jpg)
![Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308] - An intracellular stain was performed on Neuro2a cells with PKM2 Antibody NBP1-48308AF488 (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/PKM2-Antibody---BSA-Free-Flow-Cytometry-NBP1-48308-img0019.jpg)
![Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of HeLa lysates using NBP1-48308. Image courtesy of Gregg Semenza, (PMID: 21620138).](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0010.jpg)
![Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in MCF7 whole cell lysates.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0012.jpg)
![Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Expression of PKM1 and PKM2 in clinical colorectal cancer samples. The protein expression of PKM1 and PKM2 in clinical specimens of cancer tumor (T) and the adjacent normal tissues (N) is shown. PKM1 and PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S3b. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep08647) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0016.jpg)
![Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Expression profile of PKM isoforms in various cancer cell lines, control cell lines, and cells in primary culture. PKM1 and PKM2 were detected by Western blotting under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S2a. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep08647) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in HeLa cells.](http://images.novusbio.com/fullsize/PKM2-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NBP1-48308-img0008.jpg)
![Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308] - Immunohistochemical staining of normal colon tissue adjacent to tumor tissue of case 10. Results of staining with anti-PKM2 is shown. The boxed regions are enlarged in the images below. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25721733/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/PKM2 Antibody - BSA Free-Immunohistochemistry-NBP1-48308-img0020.jpg)
![Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308] - An intracellular stain was performed on HeLa cells with PKM2 Antibody NBP1-48308AF488 (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/PKM2-Antibody---BSA-Free-Flow-Cytometry-NBP1-48308-img0014.jpg)
![Simple Western: PKM2 Antibody - BSA Free [NBP1-48308] - Lane view shows a specific band for PKM2 in 0.1 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/PKM2-Antibody---BSA-Free-Simple-Western-NBP1-48308-img0013.jpg)
![Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Analysis in human cerebral cortex and liver tissues. Corresponding AQP4 RNA-seq data are presented for the same tissues.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0027.jpg "<b>Orthogonal Strategies Validation. </b>Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Analysis in human cerebral cortex and liver tissues. Corresponding AQP4 RNA-seq data are presented for the same tissues.")
![<b>Orthogonal Strategies Validation. </b>Western Blot: Aquaporin-4 Antibody [NBP1-87679] - Analysis in human brain tissue.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Western-Blot-NBP1-87679-img0017.jpg)
![Immunohistochemistry: Aquaporin-4 Antibody [NBP1-87679] - Staining of mouse cerebellum shows positivity in astrocytes, mainly in the molecular layer.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0029.jpg)
![Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Staining of human cerebellum shows moderate to strong membranous positivity in astrocytes, mainly in the molecular layer.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0030.jpg)
![Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Staining of human cerebral cortex shows moderate to strong membranous positivity in astrocytes.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0031.jpg)
![Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Staining of human liver shows no positivity in hepatocytes, as expected.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0032.jpg)
![Immunohistochemistry-Paraffin: Aquaporin-4 Antibody [NBP1-87679] - Staining of human stomach shows strong membranous positivity in glandular cells.](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Paraffin-NBP1-87679-img0033.jpg)
![Immunohistochemistry-Frozen: Aquaporin-4 Antibody [NBP1-87679] - Mouse trigeminal ganglion stained with Aquaporin-4 antibody at 1:5000. Image submitted by a verified customer review](http://images.novusbio.com/fullsize/Aquaporin-4-Antibody-Immunohistochemistry-Frozen-NBP1-87679-img0036.jpg)
![Immunocytochemistry/ Immunofluorescence: Palladin Antibody (1E6) [NBP1-25959] - C2C12 cells were transfected with shRNA targeting palladin or nontargeting shRNA as a control. Stable transfectants were submitted to a differentiation medium and followed for 5 days in culture (D0 to D5). Myogenic differentiation was assessed at the indicated time points using RNA/protein analysis and immunofluorescence staining. Immunofluorescence images of stable transfectants labeled with palladin antibody (green) and F-actin (red). Blue is DAPI-stained nuclei. Scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124762) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Knockdown-Validated-NBP1-25959-img0012.jpg "<b>Genetic Strategies Validation. </b>Immunocytochemistry/ Immunofluorescence: Palladin Antibody (1E6) [NBP1-25959] - C2C12 cells were transfected with shRNA targeting palladin or nontargeting shRNA as a control. Stable transfectants were submitted to a differentiation medium and followed for 5 days in culture (D0 to D5). Myogenic differentiation was assessed at the indicated time points using RNA/protein analysis and immunofluorescence staining. Immunofluorescence images of stable transfectants labeled with palladin antibody (green) and F-actin (red). Blue is DAPI-stained nuclei. Scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124762) licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Western Blot: Palladin Antibody (1E6) [NBP1-25959] - MDA-MB-231 lysate.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Western-Blot-NBP1-25959-img0003.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: Palladin Antibody (1E6) [NBP1-25959] - Western blots analysis of C2C12 cells labeled with a monoclonal antibody against 90-kDa palladin or a polyclonal antibody against 140- and 200-kDa palladin, MyHC, myogenin, and MLC. The blots clearly show the decrease in palladin expression concomitant with the elevation of differentiation markers. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0124762) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Knockdown-Validated-NBP1-25959-img0013.jpg)
![Immunocytochemistry/Immunofluorescence: Palladin Antibody (1E6) [NBP1-25959] - Immunofluorescence on a7r5 cells.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Immunocytochemistry-Immunofluorescence-NBP1-25959-img0005.jpg)
![Western Blot: Palladin Antibody (1E6) [NBP1-25959] - Western blot analysis of HeLa (A), A431 (B), NIH3T3 (C), and PC-12 (D) cell line extracts using Palladin antibody at 1ug/ml.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Western-Blot-NBP1-25959-img0009.jpg)
![Immunohistochemistry-Paraffin: Palladin Antibody (1E6) [NBP1-25959] - Palladin was detected in immersion fixed paraffin-embedded sections of human heart using Mouse Anti-Human Palladin (1E6) Monoclonal Antibody (Catalog # NBP1-25959) at 1:100 for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm in cardiomyocytes.](http://images.novusbio.com/fullsize/Palladin-Antibody-1E6-Immunohistochemistry-Paraffin-NBP1-25959-img0010.jpg)
![Flow Cytometry: Palladin Antibody (1E6) [NBP1-25959] - An intracellular stain was performed on HeLa cells with Palladin Antibody [1E6] NBP1-25959AF488 (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/Palladin-Antibody-1E6-Flow-Cytometry-NBP1-25959-img0011.jpg)
![Immunohistochemistry: Palladin Antibody (1E6) [NBP1-25959] - Palladin staining of paraffin-embedded patient tissues.A. IHC staining was performed using standard antigen-retrieval protocols, & counter-stained with hematoxylin. Tissue sections were stained for palladin using two palladin antibodies: polyclonal 622 & monoclonal 1E6. Palladin stain is detected with brown reaction product. In tumor sections, palladin is detected at dramatically elevated levels in the stromal fibroblasts. Note also the expanded stroma around the neoplastic cells, which is characteristic of the desmoplastic reaction. Scale bars, 200 µM. B. Quantification of immunohistochemistry results. Ten sections each of normal pancreatitis & adenocarcinoma specimens were stained with four different antibodies (622, COM, 1E6 & 4D10) & scored by two pathologists, as described in the text. Results for both ductal epithelium (left) & stroma (right) stained with various palladin antibodies are shown for normal pancreas (n = 9, blue), pancreatitis (n = 7, red) & pancreatic adenocarcinoma (n = 10, yellow). The results confirmed that palladin levels are increased in the stroma, & not the epithelial tumor cells, of the adenocarcinomas. Although palladin levels are also increased in cases of chronic pancreatitis, they do not reach the same levels as in the tumors. Compared to the polyclonal 622 & COM, the monoclonal antibodies 1E6 & 4D10 are effective at distinguishing between pancreatitis & cancer. C. Double-label immunostaining for palladin (1E6 Ab) & alpha -SMA in sections of pancreatic tumors confirms that palladin is strongly detected in a population of activated TAFs that surround the neoplastic cells. Scale bars, 200 µM. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0010347), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-25959_mouse-monoclonal-palladin-antibody-1e6-310202416173545.jpg)
![Western Blot: Palladin Antibody (1E6) [NBP1-25959] - Analysis of human palladin isoforms in pancreatic tissues.A. Human palladin isoforms. Proline-rich domains are represented by red boxes, & Ig-like domains are shown as blue boxes. The epitope recognized by the 1E6 & 4D10 antibodies is highlighted in green. The region amplified by RT-qPCR is highlighted in light blue. Isoform #1, 3 & 4 are the primary products of the palladin gene & have been detected by immunoblotting. The sequences of these isoforms are published. The sequences of isoforms #2, 5, 6, & 7 were obtained from genomic databases. “ND”: not-determined. B. Western blot analysis of pancreas samples. Small pieces of fresh tissue were snap-frozen in liquid nitrogen, ground in a chilled mortar & pestle, extracted in a detergent-containing lysis buffer, & centrifuged at 15,000×g to remove any unsolubilized particulates. The supernatant was boiled in Laemmli sample buffer & resolved by SDS-PAGE, with 15 µg protein loaded per lane. The samples were immunoblotted & probed with two anti-palladin antibodies & an antibody to GADPH (a housekeeping gene) as a control for equal loading. Lanes 1–2: normal pancreas. Lanes 3–5: primary adenocarcinoma tumors (PDA). Lane 6–7: Non-primary adenocarcinoma tumors (Non-PDA) (Lane 6: solid pseudopapillary tumor, Lane 7: neuroendocrine tumor). C. RT-qPCR. Total RNA was isolated from normal tissue (patients 1–4) & PDA tumors (patients 1–3), reverse transcribed, & subjected to RT-qPCR using gene-specific primers. Each bar represents the mean + SEM (0.06–0.35%) from three or more independent determinations. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0010347), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-25959_mouse-monoclonal-palladin-antibody-1e6-31020241616073.jpg)
![Western Blot: Palladin Antibody (1E6) [NBP1-25959] - Detection of palladin in post-surgical samples collected with 18-gauge needles.A. Samples of normal (lane 1 to 3) & pancreatic adenocarcinoma (lanes 4 to 7) were obtained from donated post-surgical organs using 18-gauge needles. Tissue samples were snap-frozen, ground, lysed & analyzed as in Figure 1. The blot was stained with both monoclonal (1E6) & polyclonal (622) palladin antibodies, & the major band (85–90 kDa) was detected by both antibodies in all tumor samples. B. Same samples (normal, lane 1–3 & PDA, lane 4–7) were analyzed for epithelial vs myofibroblast markers. The blot was stained with both, anti-E-cadherin antibody (as an epithelial cell marker) & anti-alpha SMA antibody (as a myofibroblast marker). Blots were stained for tubulin as a control for equal loading. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0010347), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-25959_mouse-monoclonal-palladin-antibody-1e6-310202416161829.jpg)
![Western Blot: Palladin Antibody (1E6) [NBP1-25959] - CAFs & NOFs are biochemically & morphologically different & CAF exosomes can also be transferred to CRC cells(A) Western blot of paired primary NOFs & CAFs for myofibroblastic markers alpha-smooth muscle actin ( alpha -SMA), fibronectin ED-A (ED-A FN1), palladin & vimentin. HSC-70 was used as an equal loading control. (B) Light microscopy of representative primary NOF & CAF cells (10x). (C) Fluorescence microscopy demonstrating phalloidin staining of F-actin filaments (green), counterstained with DAPI (blue; 40x). (D) Mean surface area & (E) intensity of phalloidin staining in a representative NOF-CAF pair. (F) Flow cytometry of DLD1 cells (control) & DLD1 cells co-cultured with CAF exosomes (exosome). The proportion of cells under the M1 region is given as a percentage. (G) Co-culture of CAF exosomes with DLD1 & SW480 cells with resultant increase in miR-199b & miR-21-5p. Data is presented as mean +/− SEM. Student's t-test (D, E) or paired t-test (F, G): *p<0.05, **p<0.01, ***p<0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29283887), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-25959_mouse-monoclonal-palladin-antibody-1e6-310202416173529.jpg)
![Immunocytochemistry/ Immunofluorescence: Palladin Antibody (1E6) [NBP1-25959] - Effect of palladin depletion on terminal differentiation of C2C12 cells.(A) Phase contrast images of stable transfectants at late stages of differentiation. Scale bar is 50 μm. (B) Representative immunofluorescence images of stable transfectants at day 5 of differentiation. Cells were labeled with palladin (red), MyHC (green), & DAPI (blue). Scale bar is 100 μm. (C) Fusion index analysis of stable transfectants at day 5 (left) & day 7 (right) of differentiation. A minimum of 4,000 nuclei were counted from random fields of each cell line. Note that palladin depletion resulted in a decrease of the fusion index at the late stage of differentiation. (D) Quantification of multinucleated myotubes throughout the course of differentiation. (E) Quantification of the number of MyHC-positive cells in stable transfectants. All error bars indicate the means ± SD of at least four independent experiments. * indicates statistically significant difference from control cells, * p<0.05, ** p<0.01, *** p<0.001 by Student’s t-test. ns = not significant. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25875253), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-25959_mouse-monoclonal-palladin-antibody-1e6-310202416171480.jpg)
![Western Blot: Mitofusin 1 Antibody (3C9) [H00055669-M04] - Analysis of MFN1 expression in HepG2.](http://images.novusbio.com/fullsize/Mitofusin-1-Antibody-3C9-Western-Blot-H00055669-M04-img0005.jpg "Western Blot: Mitofusin 1 Antibody (3C9) [H00055669-M04] - Analysis of MFN1 expression in HepG2.")
![Western Blot: Mitofusin 1 Antibody (3C9) [H00055669-M04] - MFN1 monoclonal antibody (M04), clone 3C9 Western Blot analysis of MFN1 expression in HeLa ( Cat # L013V1 ).](http://images.novusbio.com/fullsize/Mitofusin-1-Antibody-3C9-Western-Blot-H00055669-M04-img0001.jpg)
![Western Blot: Mitofusin 1 Antibody (3C9) [H00055669-M04] - Western blot analysis of MFN1 over-expressed 293 cell line, cotransfected with MFN1 Validated Chimera RNAi or non-transfected control. Blot probed with H00055669-M04. GAPDH (36.1 kDa) used as loading control.](http://images.novusbio.com/fullsize/Mitofusin-1-Antibody-3C9-Western-Blot-H00055669-M04-img0003.jpg)
![Western Blot: Mitofusin 1 Antibody (3C9) [H00055669-M04] - Analysis of MFN1 expression in transfected 293T cell line by MFN1 monoclonal antibody (M04), clone 3C9. Lane 1: MFN1 transfected lysatE (84.2 KDa). Lane 2: Non-transfected lysate.](http://images.novusbio.com/fullsize/Mitofusin-1-Antibody-3C9-Western-Blot-H00055669-M04-img0006.jpg)
![ELISA: Mitofusin 1 Antibody (3C9) [H00055669-M04] - Detection limit for recombinant GST tagged MFN1 is 0.3 ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/Mitofusin-1-Antibody-3C9-ELISA-H00055669-M04-img0007.jpg)
![Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - Inhibition of AKT decreases p53mut stability. T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 i1/4M), Nutlin3A (5 i1/4M) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected and cropped by Citeab from the following publication (AKT regulates NPM dependent ARF localization and p53mut stability in tumors. Oncotarget (2014)) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Western-Blot-NB200-111-img0012.jpg "<b>Genetic Strategies Validation. </b>Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - Inhibition of AKT decreases p53mut stability. T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 i1/4M), Nutlin3A (5 i1/4M) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected and cropped by Citeab from the following publication (AKT regulates NPM dependent ARF localization and p53mut stability in tumors. Oncotarget (2014)) licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Immunohistochemistry: p14ARF/CDKN2A Antibody [NB200-111] - Inhibition of AKT modulates p53 stability in-vivo and synergizes with ionizing radiation to inhibit tumor growth( Sections of PSN1 xenografts treated with three consecutive doses of MK-2206 (60 mg/kg). Sections of PSN1 xenografts and in-vitro PSN1 cells fixed and stained with anti-NPM (red) and anti-p14ARF (green). Image collected and cropped by Citeab from the following publication (AKT regulates NPM dependent ARF localization and p53mut stability in tumors. Oncotarget (2014)) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Immunocytochemistry-Immunofluorescence-NB200-111-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested in HeLa cells with Dylight 488 (green). Nuclei were counterstained with DAPI (blue). Tubulin was stained with alpha tubulin (red).](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Immunocytochemistry-Immunofluorescence-NB200-111-img0006.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - ARF expression levels in OCI/AML3 cells after proteasome treatment. This image was submitted via customer Review.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Western-Blot-NB200-111-img0009.jpg)
![Flow Cytometry: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested at 1:400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Flow-Cytometry-NB200-111-img0007.jpg)
![Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - Analysis of HeLa Whole Cell Lysate (NB800-PC1) using p14ARF antibody (lot C) employing ECL detection method.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Western-Blot-NB200-111-img0008.jpg)
![Immunohistochemistry-Frozen: p14ARF/CDKN2A Antibody [NB200-111] - Mouse brain section, hippocampal (10 um thickness). 3% PFA perfused. 1:1000 dilultion. Image from verified customer review. Image using the Biotin form of this antibody.](http://images.novusbio.com/fullsize/p14ARF-CDKN2A-Antibody-Immunohistochemistry-Frozen-NB200-111-img0010.jpg)
![Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT decreases p53mut stability(A) T24 cells & PSN1 cells were treated with MK-2206 (5 μM) or DMSO for 24 hrs. Cells were fixed & stained with DAPI & anti-mutant p53 (OP29 clone). (B) T24 cells were pre-treated with MK-2206 (5 μM) or DMSO for 24 hrs before the addition of fresh media containing cyclohexamide (100 μM) (CHX) in combination with MK-2206 (5 μM) or DMSO for the times indicated. Nuclear extracts were prepared from treated cells & blotted with the indicated antibodies. Quantification is relative to initiation of CHX treatment for both conditions *, MK-2206 is 40% of DMSO control but taken as 1.0 for relative assessment. (C) H1299 cells were transfected with HA-tagged-ubiquitin & mutant p53 (R175H or R248W) as indicated. Transfected cells were treated with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) for 16hrs as indicated. p53 immunoprecipitates & whole cell lysates were probed with the indicated antibodies. (D) T24 cells were transfected with non-targeting control or p14ARF siRNA & treated with DMSO, MK-2206 (5 μM) or the NPM oligomerisation inhibitor NCS348884 (4 μM) (Qi et al., 2008) as indicated. (E) T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 μM), Nutlin3A (5 μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. (F) KPC mice derived KRASG12D p53 Floxed (p53Fl), KRASG12D p53R172H ARF+/+ & KRASG12D p53R172H ARF−/− pancreatic tumor cells were treated with MK-2206 (1μM), Nutlin3A (5μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416203719.jpg)
![Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT decreases p53mut stability(A) T24 cells & PSN1 cells were treated with MK-2206 (5 μM) or DMSO for 24 hrs. Cells were fixed & stained with DAPI & anti-mutant p53 (OP29 clone). (B) T24 cells were pre-treated with MK-2206 (5 μM) or DMSO for 24 hrs before the addition of fresh media containing cyclohexamide (100 μM) (CHX) in combination with MK-2206 (5 μM) or DMSO for the times indicated. Nuclear extracts were prepared from treated cells & blotted with the indicated antibodies. Quantification is relative to initiation of CHX treatment for both conditions *, MK-2206 is 40% of DMSO control but taken as 1.0 for relative assessment. (C) H1299 cells were transfected with HA-tagged-ubiquitin & mutant p53 (R175H or R248W) as indicated. Transfected cells were treated with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) for 16hrs as indicated. p53 immunoprecipitates & whole cell lysates were probed with the indicated antibodies. (D) T24 cells were transfected with non-targeting control or p14ARF siRNA & treated with DMSO, MK-2206 (5 μM) or the NPM oligomerisation inhibitor NCS348884 (4 μM) (Qi et al., 2008) as indicated. (E) T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 μM), Nutlin3A (5 μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. (F) KPC mice derived KRASG12D p53 Floxed (p53Fl), KRASG12D p53R172H ARF+/+ & KRASG12D p53R172H ARF−/− pancreatic tumor cells were treated with MK-2206 (1μM), Nutlin3A (5μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416212372.jpg)
![Immunocytochemistry/ Immunofluorescence: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT modulates p53 stability in-vivo & synergizes with ionizing radiation to inhibit tumor growth(A) PSN1 xenografts (PSN1 cells co-injected with LTC-14 stellate cells) established in the flank of athymic nude mice were treated with MK-2206 (60 mg/kg-320 mg/kg) as indicated or beta -cyclo-dextrin (1.5 mg/ml) carrier. Xenograft tumors were lysed & lysates probed by western blot with the indicated antibodies. (B-D) Sections of PSN1 xenografts treated with three consecutive doses of MK-2206 (60 mg/kg). (B) Sections of PSN1 xenografts & in-vitro PSN1 cells fixed & stained with anti-NPM (red) & anti-p14ARF (green). (C) PSN1 xenografts treated with MK-2206 or carrier were stained with DAPI, anti-p53 (DO1) or p53mut (OP29 clone) (D) PSN1 xenografts treated with MK-2206 (60 mg/kg) or carrier were stained by immunohistochemical methods with anti-pS473-AKT, anti-phospho-S48-NPM (pS48-NPM) or p53. (E) PSN1 xenografts established in the flank of athymic nude mice were injected subcutaneously with two alternate day doses of MK-2206 (60 mg/kg) or carrier. Mice were subsequently treated with a single dose of IR (6 Gy) & tumor volumes measured regularly with callipers. Dash lines indicate tumor growth differential at 250 mm3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416203724.jpg)
![Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT decreases p53mut stability(A) T24 cells & PSN1 cells were treated with MK-2206 (5 μM) or DMSO for 24 hrs. Cells were fixed & stained with DAPI & anti-mutant p53 (OP29 clone). (B) T24 cells were pre-treated with MK-2206 (5 μM) or DMSO for 24 hrs before the addition of fresh media containing cyclohexamide (100 μM) (CHX) in combination with MK-2206 (5 μM) or DMSO for the times indicated. Nuclear extracts were prepared from treated cells & blotted with the indicated antibodies. Quantification is relative to initiation of CHX treatment for both conditions *, MK-2206 is 40% of DMSO control but taken as 1.0 for relative assessment. (C) H1299 cells were transfected with HA-tagged-ubiquitin & mutant p53 (R175H or R248W) as indicated. Transfected cells were treated with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) for 16hrs as indicated. p53 immunoprecipitates & whole cell lysates were probed with the indicated antibodies. (D) T24 cells were transfected with non-targeting control or p14ARF siRNA & treated with DMSO, MK-2206 (5 μM) or the NPM oligomerisation inhibitor NCS348884 (4 μM) (Qi et al., 2008) as indicated. (E) T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 μM), Nutlin3A (5 μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. (F) KPC mice derived KRASG12D p53 Floxed (p53Fl), KRASG12D p53R172H ARF+/+ & KRASG12D p53R172H ARF−/− pancreatic tumor cells were treated with MK-2206 (1μM), Nutlin3A (5μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416205197.jpg)
![Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT promotes enhanced MDM2 activity via the increased association between NPM & p14ARF(A) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus empty vector & pBABE expressing FLAG-tagged-NPM-WT, NPM-S48A or S48E as indicated. Immunopurification of NPM was done by pulling down with the Flag tag (middle panel) followed by elution of complexes by the Flag peptide & subsequent immunopurification of endogenous MDM2 (lower panel). (B) Nuclear immunoprecipitates of MDM2 from T24 cells treated with MK-2206 (5 μM, 24 hrs). Immunoprecipitates & lysates were blotted with the indicated antibodies. (C) T24 cells were treated with MK-2206 (5 μM) as indicated. p14ARF was immunoprecipitated from whole cell lysates & nuclear extracts & the association with NPM & MDM2 determined by western blot. Immunoprecipitates & lysates were blotted with the indicated antibodies. (D) MDM2 & (E) p53 ubiquinitation assay in H1299 cells transfected with wild type p53, HA-tagged ubiquitin & treated for 16 hrs with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) as indicated. Immunoprecipitates & whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416212370.jpg)
![Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT promotes enhanced MDM2 activity via the increased association between NPM & p14ARF(A) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus empty vector & pBABE expressing FLAG-tagged-NPM-WT, NPM-S48A or S48E as indicated. Immunopurification of NPM was done by pulling down with the Flag tag (middle panel) followed by elution of complexes by the Flag peptide & subsequent immunopurification of endogenous MDM2 (lower panel). (B) Nuclear immunoprecipitates of MDM2 from T24 cells treated with MK-2206 (5 μM, 24 hrs). Immunoprecipitates & lysates were blotted with the indicated antibodies. (C) T24 cells were treated with MK-2206 (5 μM) as indicated. p14ARF was immunoprecipitated from whole cell lysates & nuclear extracts & the association with NPM & MDM2 determined by western blot. Immunoprecipitates & lysates were blotted with the indicated antibodies. (D) MDM2 & (E) p53 ubiquinitation assay in H1299 cells transfected with wild type p53, HA-tagged ubiquitin & treated for 16 hrs with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) as indicated. Immunoprecipitates & whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-111_rabbit-polyclonal-p14arf-cdkn2a-antibody-310202416205120.jpg)