Name: Anti-Nanog antibody [EPR20694] - ChIP Grade
SKU: ab214549
Rating: 5 (3 reviews)

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.

Images

ChIP - Anti-Nanog antibody [EPR20694] - ChIP Grade (AB214549), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIC/CUT&RUN-seq	IP	ChIP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Mouse	Tested	Tested	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 5 µg	Notes -

Tested
Tested

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.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 5 µg for 25 µg chromatin	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/60	Notes -

Tested
Tested

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.

Associated Products

Select an associated product type

14 products for Alternative Product

Target data

See full target information Nanog

Function

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.

Alternative names

Ecat4, Enk, Nanog, Homeobox protein NANOG, ES cell-associated protein 4, Early embryo specific expression NK-type homeobox protein, Homeobox transcription factor Nanog

Recommended products

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.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20694
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

ChIP - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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).
Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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
Secondary
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
Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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
Secondary
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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.
Immunoprecipitation - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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.
Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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 sequencing - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
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.