Name: Anti-Nanog antibody [EPR2027(2)]
SKU: ab109250
Rating: 4 (11 reviews)

Anti-Nanog antibody [EPR2027(2)] (ab109250) is a rabbit monoclonal antibody that is used to detect Nanog in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF, ChIP. Suitable for Human samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 190 publications

Images

Immunoprecipitation - Anti-Nanog antibody [EPR2027(2)] (AB109250), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIP-seq	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/40	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 Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/70	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes Antigen retrieval is recommended. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information NANOG

Function

Transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. 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 (PubMed:25825768). 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

Homeobox protein NANOG, Homeobox transcription factor Nanog, hNanog, NANOG

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% 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: EPR2027(2)
Purification technique: Affinity purification Protein A
Specificity: 100% identities with NANOGP8
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

What is this antibody validated in?

Anti-Nanog antibody [EPR2027(2)] (ab109250) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), ChIP in Human samples.

What is the molecular weight of Nanog?

Anti-Nanog [EPR2027(2)] (ab109250) specifically detects a band for Nanog (UniProt: Q9H9S0) at a molecular weight of 35kDa.

Trusted by the scientific community

Anti-Nanog [EPR2027(2)] (ab109250) was first used in a scientific publication in 2011 and has been cited over 190 times in peer-reviewed journals.

Reviewed by scientists

Anti-Nanog [EPR2027(2)] (ab109250) has over 10 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR2027(2)] also available for your convenience:
ab109250, Alexa Fluor® 647 - ab195018, Alexa Fluor® 488 - ab196155, Carrier free - ab218524

Species reactivity
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

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

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14 product images

Immunoprecipitation - Anti-Nanog antibody [EPR2027(2)] (ab109250)
ab109250 (purified) at 1/500 dilution (1.5 ©:g/ml) immunoprecipitating Nanog in NCCIT whole cell lysate.
Lane 1 (input): NCCIT(Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10©:g
Lane 2 (+): ab109250 & NCCIT whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109250 in NCCIT whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .
All lanes: Immunoprecipitation - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Predicted band size: 34 kDa
Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Nanog Western blot staining using rabbit Anti-Nanog antibody
Nanog is highly expressed in cancer stem cells. Although some papers support the expression in undifferentiated cancer cell lines, such as HeLa (PMID: 22337995, 28092370), MDA-MB-231 (PMID: 28401007, 25919570), HepG2 (PMID: 29477378), Huh7 (PMID: 26919045), HCT 116 (PMID: 25249558, 28092370) and PANC-1 (PMID: 28703793, 25846752), ab109250 can't detect the target band in these cell lines, even at the dilution of 1:200.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/2000 dilution
Lane 1: NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Lane 3: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg
Lane 4: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 6: Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 7: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 8: PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 37 kDa
Exposure time: 30s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human seminoma tissue labelling Nanog with purified ab109250 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)
All lanes: Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/1000 dilution
All lanes: NCCIT cell lysate at 10 µg
Predicted band size: 34 kDa
Observed band size: 37 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human seminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human dysgerminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human embryonal carcinoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human normal colon tissue shows negative staining of Nanog with unpurified ab109250.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human adult kidney tissue shows negative staining of Nanog with unpurified ab109250.
Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Intracellular Flow Cytometry analysis ofNCCIT cells labelling Nanog with purified ab109250 at 1/70 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This image is courtesy of an anonymous customer review
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunocytochemistry/Immunofluorescence analysis of Human Liver cells labelling Nanog with unpurified ab109250. Cells were fixed with Paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 1% BSA for 12 hours at 4°C. Sample was incubated with primary antibody (1/500 in PBS) for 16 hour at 4°C. An Alexa Fluor^®647-conjugated Donkey anti-rabbit(1/1000) IgG polyclonal was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunocytochemistry/Immunofluorescence analysis of embryonic carcinoma cells labelling Nanog with unpurified ab109250 at 1/100.
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Immunocytochemistry/Immunofluorescence analysis of NCCIT(human pluripotent embryonal carcinoma) cells labelling Nanog with purified ab109250 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
ChIP-sequencing - Anti-Nanog antibody [EPR2027(2)] (ab109250)
Chromatin was prepared from NCCIT (Human pluripotent embryonic carcinoma cell line) cells. ChIP was performed with 10^7 NCCIT cells and 8 µg of ab109250 [[EPR2027(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.