Anti-PBRM1/BAF180 antibody [EPR25199-139] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-PBRM1/BAF180 antibody [EPR25199-139] - BSA and Azide free (AB305224)

This data was developed using ab305223, the same antibody clone in a different buffer formulation. PBRM1/BAF180 was immunoprecipitated from 0.35 mg HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate 10 µg with ab305223 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305223 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate 10 µg Lane 2 : ab305223 IP in HAP1 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305223 in HAP1 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 102 seconds

All lanes:

Immunoprecipitation - Anti-PBRM1/BAF180 antibody [EPR25199-139] (<a href='/en-us/products/primary-antibodies/pbrm1-baf180-antibody-epr25199-139-ab305223'>ab305223</a>) at 1/30 dilution

All lanes:

HAP1 (human chronic myelogenous leukemia near-haploid cell)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 205 kDa

false

Exposure time: 102s

• WB

Supplier Data

Western blot - Anti-PBRM1/BAF180 antibody [EPR25199-139] - BSA and Azide free (AB305224)

This data was developed using ab305223, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS The identity of the bands between 30 kDa and 200 kDa are unknown. The samples were run on a Bis-Tris gel. Performed under reducing conditions. False colour image of Western blot : Anti-PBRM1/BAF180 antibody (ab305223) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab305223 was shown to bind specifically to PBRM1/BAF180. A band was observed at 205 kDa in wild-type HAP1 cell lysates whereas no signal observed at this size in PBRM1/BAF180 knockout cell line. To generate this image, wild-type and PBRM1/BAF180 knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-PBRM1/BAF180 antibody [EPR25199-139] (<a href='/en-us/products/primary-antibodies/pbrm1-baf180-antibody-epr25199-139-ab305223'>ab305223</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate

Lane 2:

PBRM1/BAF180 knockout HAP1 whole cell lysate

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Observed band size: 205 kDa

false

• WB

Supplier Data

Western blot - Anti-PBRM1/BAF180 antibody [EPR25199-139] - BSA and Azide free (AB305224)

This data was developed using ab305223, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the bands between 30 kDa and 200 kDa are unknown. Exposure time : 158 seconds

All lanes:

Western blot - Anti-PBRM1/BAF180 antibody [EPR25199-139] (<a href='/en-us/products/primary-antibodies/pbrm1-baf180-antibody-epr25199-139-ab305223'>ab305223</a>) at 1/1000 dilution

Lane 1:

Hela (human cervical adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HEK-293T (human embryonic kidney epithelial cell), whole cell 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

Observed band size: 205 kDa

false

Exposure time: 158s