Anti-GRHL2 antibody [EPR24386-76]

8 product images

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  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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Predicted band size: 71 kDa

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  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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Predicted band size: 71 kDa

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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.

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  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.

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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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

• 

  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.

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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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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.
  

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: HeLa. (PMID: 25550776).