HAF109). A specific band was detected for Nanog at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using
Immunoblot Buffer Group 9." class="big_lightbox">
HAF109). A specific band was detected for Nanog at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using
Immunoblot Buffer Group 9." alt="Western blot shows lysates of BG01V human embryonic stem cells and Tera-2 human embryonic lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Nanog Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1997) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (
HAF109). A specific band was detected for Nanog at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using
Immunoblot Buffer Group 9." class="big_thumb" />
AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (
BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog #
MAG999) and DNA was purified using chelating resin solution. The
nanogpromoter was detected by standard PCR." class="big_lightbox">
AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (
BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog #
MAG999) and DNA was purified using chelating resin solution. The
nanogpromoter was detected by standard PCR." class="big_thumb" />
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox">
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in developing sperm cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox">
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in developing sperm cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei in developing CNS. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox">
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei in developing CNS. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />
HAF109). A specific band was detected for Nanog at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 9." alt="Western blot shows lysates of BG01V human embryonic stem cells and Tera-2 human embryonic lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Nanog Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1997) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Nanog at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 9." class="big_thumb" />
AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (
NL001) and counterstained with DAPI (blue). View our protocol for 
NL001) and counterstained with DAPI (blue). View our protocol for 
NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for 
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (
![Combinatorial histone hyperacetylation in SZ neurons.a Representative (repeated >3X/group) patient-specific hiPSCs, NPCs, and neurons. Left-hiPSCs express NANOG (green) and TRA-1-60 (red). DAPI (blue). Center-hiPSC neural progenitor cells (NPCs) express NESTIN (green) and SOX2 (red). DAPI (blue). Right-hiPSC neurons express S100 calcium-binding protein B (green) and the dendritic marker MAP2AB (red). DAPI (blue). Scale bar 100 μm. b Heatmap depicting LC-MS/MS data for relative enrichment values of (un)modified and acetylated histones H2AZ.1 and H2AZ.2 in hiPSCs (n = 3/group), NPC (n = 3/group), and 4-week-old neurons from SZ vs. matched controls (n = 4/group). Absolute values (% total peptide) for each peptide are provided. Fold differences between SZ vs. controls are represented (biologically independent replicates/cell-type/condition). Heatmap data represented as means, *p ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001 (two-tail Student’s t-tests performed within cell-type, SZ vs. CTRL; adjustments were not made for multiple comparisons). Please see Supplementary Data 1 for LC-MS/MS source data. Increased patterns of H2A.Z acetylation were confirmed via western blotting in c 4-week-old hiPSC neurons [n = 3 (SZ) vs. 4 (CTRL) biologically independent replicates], *p = 0.0538 (two-tail Student’s t-tests), and in d DLPFC from biologically independent postmortem SZ subjects vs. matched controls (n = 9 per group; two-tail Student’s t-tests), *p = 0.0510. Total H2A.Z and GAPDH were used as normalization controls, respectively. A.U. = arbitrary units (normalized to CTRL samples). Data are presented as averages ± SEM. Source data are provided in Source Data files. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35459277), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af1997_human-nanog-affinity-purified-polyclonal-ab-8120255534711.jpg)
![Silencing of OCT4, NANOG, and SOX2 in hEC cells. (A): Initial RNAi screen on hESC marker genes in hEC cells. esiRNA-treated samples were evaluated on the basis of cell morphology and changes in OCT4, NANOG, and SOX2 expression levels (by real-time PCR). Numbers at the bottom are array-based expression ratios of hESCs vs. universal reference RNA. The values for GDF3 and OTX2 are from an in-house platform (our unpublished data) and [34], respectively. Knock-down efficiencies were between 60 and > 90% throughout (not shown). (B): RNAi phenotypes in the OCT4, NANOG, and SOX2 knock-downs. Pictures were taken 2.5 days after esiRNA transfection. The morphology of unmanipulated or mock-treated cells was dependent on the seeding density. When plated at low density as required for esiRNA transfections the cells grew as 3D-shaped colonies rather than in monolayers. Bottom left: Growth curves of NANOG vs. GAPDH esiRNA-transfected cells. (C): Immunostaining of OCT4 protein in samples prepared as in (B). Note that the NANOG RNAi cells are OCT4 positive. (D): Western blot on day 3 RNAi and mock control samples using OCT4, NANOG, and SOX2 antibodies. GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/17506876), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af1997_human-nanog-affinity-purified-polyclonal-ab-8120255552898.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)
![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)
![Immunohistochemistry: Nanog Antibody - BSA Free [NBP1-77109] - Validation of Nanog in Rat Testis Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Nanog antibody at 2 u/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-2711202419592087.jpg "Immunohistochemistry: Nanog Antibody - BSA Free [NBP1-77109] - Validation of Nanog in Rat Testis Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Nanog antibody at 2 u/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.")
![Immunohistochemistry: Nanog Antibody - BSA Free [NBP1-77109] - Validation of Nanog in Mouse Testis.Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Nanog antibody at 2 u/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-2711202420312999.jpg)
![Immunohistochemistry: Nanog Antibody - BSA Free [NBP1-77109] - Validation of Nanog in Human Testis.Immunohistochemical analysis of paraffin-embedded human testis tissue using anti-Nanog antibody at 5 u/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-271120241932686.jpg)
![Immunocytochemistry/ Immunofluorescence: Nanog Antibody - BSA Free [NBP1-77109] - Validation of Nanog in HeLa Cells.Immunofluorescent analysis of PFA-fixed HeLa cells labeling Nanog with at 20 u/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-2711202419553033.jpg)
![Western Blot: Nanog Antibody - BSA Free [NBP1-77109] - Validation in Mouse and Rat Testis.Antibodies: Nanog, 2 u/mL, 1h incubation at RT in 8% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-271120241923639.gif)
![Western Blot: Nanog Antibody - BSA Free [NBP1-77109] - Validation in HeLa CellsLoading: 15 u of lysate. Antibodies: Nanog, 2 u/mL, 1 h incubation at RT in 8% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.](http://images.novusbio.com/fullsize/nbp1-77109_rabbit-polyclonal-nanog-antibody-2711202419463442.gif)
![Western Blot: Nanog Antibody (5A10) [NBP1-04320] - The cell lysates of NIH3T3 (35ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human Nanog (1:500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.](http://images.novusbio.com/fullsize/Nanog-Antibody-5A10-Western-Blot-NBP1-04320-img0001.jpg "Western Blot: Nanog Antibody (5A10) [NBP1-04320] - The cell lysates of NIH3T3 (35ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human Nanog (1:500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.")
![Flow Cytometry: Nanog Antibody (5A10) [NBP1-04320] - Flow cytometry analysis of Nanog in Hep3B cell line, staining at 2-5ug for 1x106cells. The secondary antibody used goat anti-mouse IgG Alexa fluor 488 conjugate.](http://images.novusbio.com/fullsize/Nanog-Antibody-5A10-Flow-Cytometry-NBP1-04320-img0003.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (5A10) [NBP1-04320] - Human seminoma tissue were incubated with anti-human Nanog (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodiumcitrate buffer and detected using Diaminobenzidine (DAB)](http://images.novusbio.com/fullsize/Nanog-Antibody-5A10-Immunohistochemistry-Paraffin-NBP1-04320-img0004.jpg)
![Immunocytochemistry/ Immunofluorescence: Nanog Antibody (5A10) - BSA Free [NBP1-04320] - Representative IF IHC-stained sections of MDBMSCC demonstrating the expression of pSTAT3 [(A), red] & EMA [(A), green] by cells within the tumor nests. There was a CSC subpopulation remonstrating nuclear co-expression of STAT3 [(B), red] & CD34 [(B), green], appearing as orange, on the endothelium of the microvessels within the stroma; & another subpopulation staining only positively for pSTAT3 within the stroma [(B), red]. Nuclear expression of NANOG [(C), red] was demonstrated on the endothelium of the microvessels which expressed CD34 [(C), green] within stroma. The NANOG+ cells [(C), red] that do not express CD34 were seen within the tumor nests & the stroma. SOX2 [(D), red] was also expressed by cells within tumor nests & the stroma, & the endothelium of the microvessels expressing CD34 [(D), green]. Nuclear expression of both SOX2 [(E), red] & SALL4 [(E), green], appearing as orange, was seen on the cells within the tumor nests & the stroma. Expression of both SOX2 [(F), red] & OCT4 [(F), green], appearing as orange, was seen on cells within the tumor nests & the stroma, & the endothelium of the microvessels within the stroma. pSTAT3 [(G), red] & membranous staining CD44 [(G), green] were co-expressed by cells within the tumor nests. Scale bars: 20 μm. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/Article/10.3389/fsurg.2016.00046/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-04320_mouse-nanog-mab-5a10-bsa-free-310202416165642.jpg)
![Western Blot: Nanog Antibody [NBP2-24941] - Analysis of Nanog in (A) HeLa and (B) NIH 3T3 cell lysate using NBP2-24941 at 2 ug/ml.](http://images.novusbio.com/fullsize/Nanog-Antibody-Western-Blot-NBP2-24941-img0002.jpg "Western Blot: Nanog Antibody [NBP2-24941] - Analysis of Nanog in (A) HeLa and (B) NIH 3T3 cell lysate using NBP2-24941 at 2 ug/ml.")
![Immunocytochemistry/Immunofluorescence: Nanog Antibody [NBP2-24941] - Nanog antibody was tested in Ntera2 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red). Image objective 40x. An antibody dilution of 1:10 was used.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-24941-img0005.jpg)
![Immunohistochemistry: Nanog Antibody [NBP2-24941] - Immunodetection of pluripotency markers in uterine tissues from control cows & from cows with adenomyosis. a, b – haematoxylin & eosin stained slides of control & adenomyotic uterus, respectively; c, d – NANOG immunodetection in control & adenomyotic tissue, respectively; e, f – immunolocalisation of OCT4 in normal & adenomyotic tissue, respectively; g, h – SOX2 immunodetection in control & adenomyotic tissue, respectively; i, j, k – no Ab, negative controls for NANOG, OCT4 & SOX2, respectively. Unspecific IgG controls (pictures not shown) served similar pictures as no Ab control. Arrows indicate the most intense histochemical reactions; dotted line indicate endometrial-myometrial border; e – endometrium, m – myometrium, es – endometrial stroma, eg – endometrial gland, a – adenomyotic lesion, v – vessel. Scale bars: 20 μm Image collected & cropped by CiteAb from the following publication (//www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-24941_rabbit-polyclonal-nanog-antibody-310202416121911.jpg)
![Western Blot: Nanog Antibody [NB110-40660] - Staining of Human Ovary lysate (35 ug protein in RIPA buffer). Antibody at 0.03 ug/mL. Primary incubation was 1 hour. Detected by chemiluminescence.](http://images.novusbio.com/fullsize/Nanog-Antibody-Western-Blot-NB110-40660-img0002.jpg "Western Blot: Nanog Antibody [NB110-40660] - Staining of Human Ovary lysate (35 ug protein in RIPA buffer). Antibody at 0.03 ug/mL. Primary incubation was 1 hour. Detected by chemiluminescence.")
![Immunocytochemistry/Immunofluorescence: Nanog Antibody [NB110-40660] - Staining (green) parts of a colony of induced pluriform stem cells derived from Human Keratinocytes. Antibody at 5 ug/mL. Data kindly provided by CMRB, Center of Regenerative Medicine in Barcelona, Spain.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunocytochemistry-Immunofluorescence-NB110-40660-img0001.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody [NBP2-13177] - Staining of Nanog in mouse testis using DAB with hematoxylin counterstain.](http://images.novusbio.com/fullsize/Nanog-Antibody-Immunohistochemistry-Paraffin-NBP2-13177-img0002.jpg "Immunohistochemistry-Paraffin: Nanog Antibody [NBP2-13177] - Staining of Nanog in mouse testis using DAB with hematoxylin counterstain.")
![Immunocytochemistry/Immunofluorescence: Nanog Antibody (1E6C4) [NBP1-47427] - Confocal immunofluorescence analysis of NTERA-2 cells (left) and HeLa cells (right) using anti-Nanog Mouse mAb (green). Red: Actin filaments have been labeled with DY-554 phalloidin.](http://images.novusbio.com/fullsize/Nanog-Antibody-1E6C4-Immunocytochemistry-Immunofluorescence-NBP1-47427-img0003.jpg "Immunocytochemistry/Immunofluorescence: Nanog Antibody (1E6C4) [NBP1-47427] - Confocal immunofluorescence analysis of NTERA-2 cells (left) and HeLa cells (right) using anti-Nanog Mouse mAb (green). Red: Actin filaments have been labeled with DY-554 phalloidin.")
![Western Blot: Nanog Antibody (1E6C4) [NBP1-47427] - Western blot analysis using anti-Nanog monoclonal antibody against NTERA-2 cell lysate (lane 2).](http://images.novusbio.com/fullsize/Nanog-Antibody-1E6C4-Western-Blot-NBP1-47427-img0004.jpg)
![Western Blot: Nanog Antibody (1E6C4) [NBP1-47427] - Whole cell protein from NTERA-2 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 mg/ml anti-Nanog in 1% milk. Protein was detected with an anti-mouse HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/Nanog-Antibody-1E6C4-Western-Blot-NBP1-47427-img0005.jpg)
![Western Blot: Nanog Antibody (CL5810) [NBP2-61429] - Analysis in human cell line NTERA-2.](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5810-Western-Blot-NBP2-61429-img0006.jpg "Western Blot: Nanog Antibody (CL5810) [NBP2-61429] - Analysis in human cell line NTERA-2.")
![Immunocytochemistry/Immunofluorescence: Nanog Antibody (CL5810) [NBP2-61429] - Staining of NTERA-2 cells using the Anti-NANOG monoclonal antibody, showing specific staining in the nucleoplasm in green. Microtubule and nuclear probes are visualized in red and blue, respectively (where available).](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5810-Immunocytochemistry-Immunofluorescence-NBP2-61429-img0001.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5810) [NBP2-61429] - Staining of human testis shows absence of positivity in seminiferous ducts as expected (negative control).](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5810-Immunohistochemistry-Paraffin-NBP2-61429-img0004.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5810) [NBP2-61429] - Staining of human testis cancer shows strong nuclear immunoreactivity in tumor cells.](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5810-Immunohistochemistry-Paraffin-NBP2-61429-img0002.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5810) [NBP2-61429] - Staining of human renal cancer shows absence of positivity in tumor cells as expected (negative control).](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5810-Immunohistochemistry-Paraffin-NBP2-61429-img0003.jpg)
![Western Blot: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of 60 ug of various whole cell lysates and 10ul of PageRuler Prestained Protein Ladder.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Western-Blot-NBP2-22632-img0006.jpg "Western Blot: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of 60 ug of various whole cell lysates and 10ul of PageRuler Prestained Protein Ladder.")
![Immunocytochemistry/Immunofluorescence: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of Nanog (green) in H9 embryonic stem cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Nanog monoclonal antibody at a dilution of 1:200 overnight at 4C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Immunocytochemistry-Immunofluorescence-NBP2-22632-img0005.jpg)
![<b>Biological Strategies Validation. </b>Flow Cytometry: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of Nanog (blue histogram) on H9 embryonic stem cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37C. Cells were incubated with a Nanog monoclonal antibody or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500ul of FACS buffer containing 10ul of 4% paraformaldehyde.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Flow-Cytometry-NBP2-22632-img0002.jpg)
![Western Blot: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of 60 ug of NCCIT nuclear and cytoplasmic fractions lysates and 10ul of PageRuler Prestained Protein Ladder](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Western-Blot-NBP2-22632-img0007.jpg)
![Immunocytochemistry/Immunofluorescence: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of Nanog (green) in NTERA-2 and HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a Nanog monoclonal antibody at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody. F-Actin (red) was stained with DyLight-554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Immunocytochemistry-Immunofluorescence-NBP2-22632-img0003.jpg)
![Immunocytochemistry/Immunofluorescence: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of Nanog (green) in HEL 11.4 induced IPS cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Nanog monoclonal antibody at a dilution of 1:200 overnight at 4C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Immunocytochemistry-Immunofluorescence-NBP2-22632-img0004.jpg)
![<b>Biological Strategies Validation. </b>Flow Cytometry: Nanog Antibody (23D2-3C6) [NBP2-22632] - Analysis of Nanog (blue histogram) on HEL 11.4 induced IPS cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37C. Cells were incubated with a Nanog monoclonal antibody or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500ul of FACS buffer containing 10ul of 4% paraformaldehyde.](http://images.novusbio.com/fullsize/Nanog-Antibody-23D2-3C6-Flow-Cytometry-NBP2-22632-img0001.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5803) [NBP2-61428] - Staining of human testis shows absence of positivity as expected (negative control).](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5803-Immunohistochemistry-Paraffin-NBP2-61428-img0004.jpg "Immunohistochemistry-Paraffin: Nanog Antibody (CL5803) [NBP2-61428] - Staining of human testis shows absence of positivity as expected (negative control).")
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5803) [NBP2-61428] - Staining of human testis cancer shows strong nuclear immunoreactivity in tumor cells.](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5803-Immunohistochemistry-Paraffin-NBP2-61428-img0002.jpg)
![Immunohistochemistry-Paraffin: Nanog Antibody (CL5803) [NBP2-61428] - Staining of human bladder cancer shows absence of positivity in tumor cells as expected (negative control).](http://images.novusbio.com/fullsize/Nanog-Antibody-CL5803-Immunohistochemistry-Paraffin-NBP2-61428-img0003.jpg)
