Rabbit Recombinant Monoclonal NANOG antibody. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse samples. Cited in 6 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
ChIC/CUT&RUN-seq | IP | ChIP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
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Mouse | Tested | Tested | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Mouse | Dilution info 5 µg | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 5 µg for 25 µg chromatin | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/60 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal (PubMed:25825768). Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes. Acts as a transcriptional activator or repressor. Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3'. Binds to the POU5F1/OCT4 promoter (By similarity). Able to autorepress its expression in differentiating (ES) cells: binds to its own promoter following interaction with ZNF281/ZFP281, leading to recruitment of the NuRD complex and subsequent repression of expression. When overexpressed, promotes cells to enter into S phase and proliferation.
Ecat4, Enk, Nanog, Homeobox protein NANOG, ES cell-associated protein 4, Early embryo specific expression NK-type homeobox protein, Homeobox transcription factor Nanog
Rabbit Recombinant Monoclonal NANOG antibody. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse samples. Cited in 6 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Nanog also known as Nanog homeobox is a transcription factor playing an important role in maintaining the pluripotency of embryonic stem cells. The molecular weight of Nanog is approximately 35 kDa. It is found mainly in the inner cell mass of the blastocyst but it also expresses in embryonic stem cells and some adult stem cell populations. Nanog acts by binding to DNA in a sequence-specific manner to regulate gene expression essential for stem cell self-renewal and pluripotency.
Nanog plays a central role in stem cell biology operating as part of a complex regulatory network. Nanog interacts with other important transcription factors like Oct4 and Sox2 forming a core pluripotency network. This network suppresses the differentiation of stem cells by regulating the expression of genes involved in cell fate decisions. By modulating the expression of different pathways Nanog ensures the maintenance of an undifferentiated state in stem cells.
Nanog is deeply embedded in the Wnt/β-Catenin and TGF-β signaling pathways important for stem cell maintenance and differentiation. In the Wnt/β-Catenin pathway Nanog works alongside proteins like β-catenin to drive the expression of genes that promote self-renewal. Meanwhile in the TGF-β pathway Nanog acts with proteins such as Smad2/3 to balance pluripotency and differentiation signals. These pathways critically support the complex network that sustains stem cell identity and function.
Nanog's expression relates closely to its role in various cancers such as glioblastoma and colorectal cancer. Abnormally high levels of Nanog contribute to the tumorigenicity of cancer cells by maintaining their self-renewal and undifferentiated state similar to its role in stem cells. Nanog interacts with proteins like p53 known for its tumor suppressor functions often leading to challenges in cancer treatment. Understanding Nanog's influence in these pathways provides valuable insights into therapeutic targets for combating cancer stem cell resilience.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
Chromatin was prepared from F9 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25 μg of chromatin, 5μg of ab214549 (red), and 20 μl of protein A/G sepharose beads slurry (10 μl of sepharose A beads + 10 μl of sepharose G beads). Then 5 μg of rabbit normal IgG was added to the control beads (grey). The immunoprecipitated DNA was quantified by real time PCR (SYBR green chemistry).
Blocking/Dilution buffer: 5% NFDM/TBST.
The multiple bands observed are consistent with the literature (PMID: 24936455). Negative control: NIH/3T3 (PMID: 12787505).
All lanes: Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549) at 1/1000 dilution
Lane 1: F9 (Mouse embryonic testicular cancer cell line) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 34 kDa
Observed band size: 29-42 kDa
Exposure time: 70s
Blocking/Dilution buffer: 5% NFDM/TBST.
The multiple bands observed are consistent with the literature (PMID: 24936455).
All lanes: Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549) at 1/1000 dilution
All lanes: ES-D3 (mouse embryonic multipotent stem cell line) whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 34 kDa
Observed band size: 29-42 kDa
Exposure time: 3min
Immunohistochemical analysis of paraffin-embedded mouse E14.5 testis tissue labeling Nanog with ab214549 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Mainly nuclear staining is observed in testis of mouse embryo E14.5 (PMID: 15939376; PMID: 12787505). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded adult mouse testis tissue labeling Nanog with ab214549 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Negative tissue: No staining on adult mouse testis is observed(PMID: 12787505; PMID: 15939376).
Counter stained with Hematoxylin.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nanog was immunoprecipitated from 0.35 mg of F9 (mouse embryonic testicular cancer cell line) whole cell lysate with ab214549 at 1/1000 dilution. Western blot was performed from the immunoprecipitate using ab214549 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5,000 dilution
Lane 1: F9 whole cell lysate 10 μg (input)
Lane 2: ab214549 IP in F9 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab214549 in F9 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
The multiple bands observed are consistent with the literature (PMID: 24936455).
All lanes: Immunoprecipitation - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
Predicted band size: 34 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeablized F9 (mouse embryonic testicular cancer cell line) and NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Nanog with ab214549 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in F9 cell line. Negative control: NIH/3T3 (PMID: 17352742).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Intracellular flow cytometric analysis of 4 % paraformaldehyde-fixed, 90% methanol permeabilized F9 (mouse embryonic testicular cancer cell line, Right) and NIH/3T3 (mouse embryo fibroblast cell line, Left) cells labeling Nanog with ab214549 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: NIH/3T3 cell line.
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 F9 (Mouse embryonic testicular cancer cell line) cells and 5µg of ab214549 [EPR20694]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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