Anti-Nanog 抗体 [EPR2027(2)]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (AB109250)

Immunocytochemistry/Immunofluorescence analysis of embryonic carcinoma cells labelling Nanog with unpurified ab109250 at 1/100.

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• IHC-P

Unknown

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.

• ICC/IF

AbReview32299****

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.

This image is courtesy of an anonymous Abreview

• IHC-P

Supplier Data

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.

• ICC/IF

Unknown

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).

• Flow Cyt (Intra)

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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.

• IHC-P

Unknown

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.

• IP

Unknown

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 (ab172730) instead of ab109250 in NCCIT whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (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

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• ChIP-seq

Supplier Data

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.

• WB

Lab

Western blot - Anti-Nanog antibody [EPR2027(2)] (AB109250)

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 34 kDa

Observed band size: 37 kDa

false

Exposure time: 30s

• WB

Unknown

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

false