Anti-PBRM1/BAF180 antibody [EPR15860]

6 product images

• 

  Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: PBRM1/BAF180 knockout HAP1 cell lysate (20 μg)
  Lane 3: Human kidney tissue lysate (20 μg)
  Lane 4: HeLa cell lysate (20 μg)
  

  Lanes 1 - 4: Merged signal (red and green). Green - ab196022 observed at 193 kDa. Red - loading control, ab18058, observed at 124 kDa.
  
  ab196022 was shown to recognize PBRM1/BAF180 when PBRM1/BAF180 knockout samples were used, along with additional cross-reactive bands. Wild-type and PBRM1/BAF180 knockout samples were subjected to SDS-PAGE. ab196022 and ab18058 (loading control to Vinculin) were diluted 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  Predicted band size: 193 kDa

• 

  Flow Cytometry (Intracellular) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  ab196022 staining PBRM1/BAF180 in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.
  

  Isoytype control: Rabbit monoclonal IgG (Black)

  Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

• 

  Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  5% NFDM/TBST: Blocking and diluting buffer.

  All lanes: Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022) at 1/10000 dilution

  All lanes: Human fetal kidney lysate at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 193 kDa

  Observed band size: 193 kDa, 35 kDa

  Exposure time: 3min

• 

  Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  5% NFDM/TBST: Blocking and diluting buffer.

  All lanes: Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022) at 1/10000 dilution

  All lanes: HeLa (Human epithelial cells from cervix adenocarcinoma) nuclear lysate at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 193 kDa

  Observed band size: 193 kDa

  Exposure time: 15s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PBRM1/BAF180 using ab196022 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on Human kidney tissue is observed. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PBRM1/BAF180 with ab196022 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). The nuclear counter stain is DAPI (blue).
  

  Nuclear staining on HeLa cell line is observed.

  -ve control - Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution. The nuclear counter stain is DAPI (blue).