Antibodies, Proteins, Kits and Reagents for Life Science

378 595 ₸ Product size

100 µl 378 595 ₸

Order now and get it on Tuesday March 02, 2021

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, IP
Reacts with: Human

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 & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

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

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.

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

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

Flow Cytometry analysis of NCCIT 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).
Anti-Nanog antibody [EPR2027(2)] (ab109250)

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