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

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Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2027(2)] to Nanog
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIP-sequencing, IP
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Product name

Anti-Nanog antibody [EPR2027(2)]
See all Nanog primary antibodies
Description

Rabbit monoclonal [EPR2027(2)] to Nanog
Host species

Rabbit
Specificity

100% identities with NANOGP8
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIP-sequencing, IPmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: NCCIT cell lysate. IHC-P: Human seminoma tissue, Human dysgerminoma tissue and Human embryonal carcinoma tissue. ICC/IF: Human embryonic carcinoma and Human liver cell lines. IP: NCCIT whole cell lysate. ChIP-seq: NCCIT cells.
General notes
Please note that Nanog is expressed variably in different tissues and that optimisation may be required depending on the tissue used for the experiment.
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR2027(2)
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Nanog protein (ab134886)
Related Products
- Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109250 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(4)	1/1000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
IHC-P	(3)	1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Antigen retrieval is recommended.
ICC/IF	(4)	1/100 - 1/250.
Flow Cyt (Intra)		1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ChIP-sequencing		Use at an assay dependent concentration.
IP		1/40.

Notes
WB 1/1000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Antigen retrieval is recommended.
ICC/IF 1/100 - 1/250.
Flow Cyt (Intra) 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ChIP-sequencing Use at an assay dependent concentration.
IP 1/40.

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 (By similarity). Acts as a transcriptional activator or repressor (By similarity). Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (By similarity). When overexpressed, promotes cells to enter into S phase and proliferation.
Tissue specificity

Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.
Sequence similarities

Belongs to the Nanog homeobox family.
Contains 1 homeobox DNA-binding domain.
Developmental stage

Expressed in embryonic stem (ES) and carcinoma (EC) cells. Expressed in inner cell mass (ICM) of the blastocyst and gonocytes between 14 and 19 weeks of gestation (at protein level). Not expressed in oocytes, unfertilized oocytes, 2-16 cell embryos and early morula (at protein level). Expressed in embryonic stem cells (ES). Expression decreases with ES differentiation.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q9H9S0 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 100293888 Human
- Entrez Gene: 79923 Human
- Omim: 607937 Human
- SwissProt: Q9H9S0 Human
- Unigene: 635882 Human
Alternative names
- Embryonic stem cell specific homeobox protein (Nanog) antibody
- ENK antibody
- FLJ12581 antibody
- hNanog antibody
- Homeobox protein NANOG antibody
- Homeobox transcription factor Nanog antibody
- homeobox transcription factor Nanog-delta 48 antibody
- NANOG antibody
- Nanog homeobox antibody
- NANOG_HUMAN antibody
see all

Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

All lanes : Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/2000 dilution

Lane 1 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 6 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 7 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 8 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

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.
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. 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.
Immunoprecipitation - Anti-Nanog antibody [EPR2027(2)] (ab109250)

ab109250 (purified) at 1/40 dilution (1.5 µg/ml) immunoprecipitating Nanog in NCCIT whole cell lysate.

Lane 1 (input): NCCIT(Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab109250 & NCCIT whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109250 in NCCIT whole cell lysate

For western blotting, ab109250 at 1/500 dilution (1.5 µg/ml) VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM /TBST.
Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/1000 dilution (unpurified) + NCCIT cell lysate at 10 µg

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
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.
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)This image is courtesy of an anonymous Abreview

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.
See Abreview
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. 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. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, 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.
Additional screenshots of mapped reads can be downloaded here.
Anti-Nanog antibody [EPR2027(2)] (ab109250)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols
Protocol Booklet

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab109250? Please let us know so that we can cite the reference in this datasheet.

ab109250 has been referenced in 112 publications.

Zhao H et al. Gli affects the stemness and prognosis of epithelial ovarian cancer via homeobox protein NANOG. Mol Med Rep 23:N/A (2021). PubMed: 33313950
Sah SK et al. Generation of Keratinocytes from Human Induced Pluripotent Stem Cells Under Defined Culture Conditions. Cell Reprogram 23:1-13 (2021). PubMed: 33373529
Li X et al. HES1 promotes breast cancer stem cells by elevating Slug in triple-negative breast cancer. Int J Biol Sci 17:247-258 (2021). PubMed: 33390847
Yang X et al. Downregulation of hsa_circ_0026123 suppresses ovarian cancer cell metastasis and proliferation through the miR-124-3p/EZH2 signaling pathway. Int J Mol Med 47:668-676 (2021). PubMed: 33416105
Lin J et al. Hypoxia increases the expression of stem cell markers in human osteosarcoma cells. Oncol Lett 21:217 (2021). PubMed: 33613706

View all Publications for this product

Customer reviews and Q&As

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1-10 of 12 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (DU145 Prostate cancer cell line)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

DU145 Prostate cancer cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 10 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (human pluripotent stem cells)

Permeabilization

Yes - Triton

Specification

human pluripotent stem cells

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 14 2019

Application

Western blot

Sample

Human Cell lysate - whole cell (patient derived iPSC)

Gel Running Conditions

Reduced Denaturing

Loading amount

10 µg

Specification

patient derived iPSC

Blocking step

Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 05 2015

Application

Immunocytochemistry/ Immunofluorescence

Sample

Horse Cell (testis tissues)

Specification

testis tissues

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Heejun Jung

Verified customer

Submitted Aug 19 2015

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (iPSC)

Permeabilization

Yes - 0.25% TritonX

Specification

iPSC

Blocking step

BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3%

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 22 2015

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (seminoma tumor)

Specification

seminoma tumor

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Leica Bond Epitope Retrieval 2

Permeabilization

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 02 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Marmoset (common) Tissue sections (embryonic testis E92)

Specification

embryonic testis E92

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution

Permeabilization

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. 煜欽林

Verified customer

Submitted Feb 08 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (SW480 cells, HCT116 cells)

Loading amount

25 µg

Specification

SW480 cells, HCT116 cells

Gel Running Conditions

Reduced Denaturing (4-12%)

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 17 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (liver cancer)

Loading amount

20 µg

Specification

liver cancer

Gel Running Conditions

Reduced Denaturing (10%gel)

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 18 2012

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (liver cancer)

Specification

liver cancer

Fixative

Paraformaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA

Permeabilization

Yes - Triton-100

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 15 2012

1-10 of 12 Abreviews or Q&A

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (112)

Customer reviews and Q&As

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nanog antibody [EPR2027(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nanog antibody [EPR2027(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nanog antibody [EPR2027(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nanog antibody [EPR2027(2)]