Anti-Ctip1/BCL-11A antibody [EPR14943-44]

8 product images

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  Immunoprecipitation - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  ab191401 (purified) at 1/500 dilution (2.1 ©:g/ml) immunoprecipitating Ctip1 / BCL-11A in Raji whole cell lysate.
  Lane 1 (input): Raji(Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10©:g
  Lane 2 (+): ab191401 & Raji whole cell lysate
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab191401 in Raji whole cell lysate
  For western blotting, VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/1000 dilution.
  Blocking and diluting buffer: 5% NFDM /TBST .

  All lanes: Immunoprecipitation - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Predicted band size: 91 kDa

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  Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Lane 1: Wild type HAP1 whole cell lysate (20 μg)
  Lane 2: Empty
  Lane 3: BCL11A (Ctip1) knockout HAP1 whole cell lysate (20 μg)
  Lane 4: Raji whole cell lysate (20 μg)
  

  Lanes 1 - 4: Merged signal (red and green). Green - ab191401 observed at 91 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

  ab191401 was shown to recognize BCL11A (Ctip1) in wild type cells as signal was lost at the expected MW in BCL11A (Ctip1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BCL11A (Ctip1) knockout samples were subjected to SDS-PAGE. ab191401 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Predicted band size: 91 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Immunohistochemical analysis of paraffin-embedded, rat spleen tissue labeling Ctip1/BCL-11A with ab191401 at a 1/100 dilution (19 μg/ml). Counter stained with hematoxylin. In the negative control PBS was used instead of primary antibody.
  

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunocytochemistry/ Immunofluorescence - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Immunofluorescence analysis of, paraformaldehyde-fixed, rat C6 cells labeling Ctip1/BCL-11A with ab191401 at a 1/450 dilution (4 ug/ml). As secondary antibody goat anti-rabbit IgG (Alexa Fluor®488) Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 was used at a 1/200 dilution. In blue DAPI staining. In the negative controls cells were treated with anti-BCL11A at a 1/450 dilution as primary antibody and goat anti-mouse IgG (Alexa Fluor®594) at a 1/400 dilution as secondary antibody.

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  Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  All lanes: Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401) at 1/50000 dilution

  Lane 1: Raji cell lysate at 20 µg

  Lane 2: Jurkat cell lysate at 20 µg

  Secondary

  All lanes: goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

  Developed using the ECL technique.

  Predicted band size: 91 kDa

  Observed band size: 120 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Immunohistochemical analysis of paraffin-embedded, Human tonsil tissue labeling Ctip1/BCL-11A with ab191401 at a 1/100 dilution (19 μg/ml). Counter stained with hematoxylin. In the negative control PBS was used instead of primary antibody.
  

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  All lanes: Western blot - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401) at 1/50000 dilution

  All lanes: 293 cell lysate at 10 µg

  Secondary

  All lanes: goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

  Developed using the ECL technique.

  Predicted band size: 91 kDa

  Observed band size: 120 kDa

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  Flow Cytometry (Intracellular) - Anti-Ctip1/BCL-11A antibody [EPR14943-44] (ab191401)

  Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling Ctip1/BCL-11A with purified ab191401 at 1/200 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor^® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.