Anti-PBRM1/BAF180 antibody [EPR15860]

• Flow Cyt (Intra)

Lab

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)

• ICC/IF

Supplier Data

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) (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) (ab150077) secondary antibody at 1/500 dilution. The nuclear counter stain is DAPI (blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBRM1/BAF180 antibody [EPR15860] (AB196022)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PBRM1/BAF180 with ab196022 at 1/200 dilution.

Nuclear staining on human kidney

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

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

false

Exposure time: 15s

• WB

Supplier Data

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

false

Exposure time: 3min

• WB

Lab

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 (ab216773) and 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

false