Anti-GRHL2 antibody [EPR24386-76]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling GRHL2 with ab271023 at 1/50 (10.52 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green) Confocal image showing nuclear staining in MCF7 cell line. Negative control : HeLa (PMID : 25550776) is observed.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labelling GRHL2 with ab271023 at 1/10000 (0.053 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human skin. The section was incubated with ab271023 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell, Left) / MCF7 (Human breast adenocarcinoma epithelial cell, Right) cells labelling GRHL2 with ab271023 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control : HeLa. (PMID : 25550776).

• IP

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Immunoprecipitation - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

GRHL2 was immunoprecipitated from 0.35 mg MDA-MB-468 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab271023 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271023 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MDA-MB-468 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab271023 IP in MDA-MB-468 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271023 in MDA-MB-468 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-GRHL2 antibody [EPR24386-76] (ab271023)

Predicted band size: 71 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labelling GRHL2 with ab271023 at 1/10000 (0.053 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse skin. The section was incubated with ab271023 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Immunohistochemical analysis of paraffin-embedded Rat skin tissue labelling GRHL2 with ab271023 at 1/10000 (0.053 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat skin. The section was incubated with ab271023 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• WB

Lab

Western blot - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight observed is consistent with what has been described in the literature (PMID : 23814079 ). The lower bands would be different GRHL2 variants.

Negative control : MDA-MB-231 (PMID : 23814079), HeLa (PMID : 25550776 ).

Exposure time : Lanes 1-6 : 26 seconds;

Lane 7 : 70 seconds.

All lanes:

Western blot - Anti-GRHL2 antibody [EPR24386-76] (ab271023) at 1/1000 dilution

Lane 1:

MDA-MB-468 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 6:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Lane 7:

Human placenta tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 71 kDa

false

• WB

Lab

Western blot - Anti-GRHL2 antibody [EPR24386-76] (AB271023)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : NIH/3T3 (PMID : 23814079).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-GRHL2 antibody [EPR24386-76] (ab271023) at 1/1000 dilution

Lane 1:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

Mouse placenta tissue lysate at 20 µg

Lane 4:

Rat placenta tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 71 kDa

false