Anti-Bcl10 antibody [EP606Y]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Bcl10 antibody [EP606Y] (AB33905)

Intracellular Flow Cytometry analysis of Raji (human Burkitt's lymphoma) cells labeling Bcl10 (red) with ab33905 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Bcl10 antibody [EP606Y] (AB33905)

Immunofluorescence staining of Raji cells with purified ab33905 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4 % PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab33905 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl10 antibody [EP606Y] (AB33905)

Unpurified ab33905, at a 1/100 dilution, staining human hepatocellular carcinoma by Immunohistochemistry, Paraffin embedded tissue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Bcl10 antibody [EP606Y] (AB33905)

Unpurified ab33905, staining HeLa cells by Immunofluorescent.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl10 antibody [EP606Y] (AB33905)

Immunohistochemical staining of paraffin embedded human lung carcinoma with purified ab33905 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IP

Unknown

Immunoprecipitation - Anti-Bcl10 antibody [EP606Y] (AB33905)

ab33905 (purified) at 1/20 immunoprecipitating Bcl10 in 10 μg Ramos cell lysate (Lanes 1 and 2, observed at 32 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used for detection (1/10 000). Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Bcl10 antibody [EP606Y] (ab33905)

Predicted band size: 26 kDa

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• WB

Lab

Western blot - Anti-Bcl10 antibody [EP606Y] (AB33905)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Bcl10 antibody [EP606Y] (ab33905) at 1/5000 dilution

Lane 1:

Raji cell lysate at 20 µg

Lane 2:

Ramos cell lysate at 20 µg

Lane 3:

HuT-78 cell lysate at 20 µg

Lane 4:

HeLa 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 1/20000 dilution

Predicted band size: 26 kDa

Observed band size: 32 kDa

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• WB

Lab

Western blot - Anti-Bcl10 antibody [EP606Y] (AB33905)

Lanes 1-4 : Merged signal (red and green). Green - ab33905 observed at 32 kDa. Red - loading control, ab7291 observed at 52 kDa.

ab33905 Anti-Bcl10 antibody [EP606Y] was shown to specifically react with Bcl10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261797 (knockout cell lysate ab257144) was used. Wild-type and Bcl10 knockout samples were subjected to SDS-PAGE. ab33905 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bcl10 antibody [EP606Y] (ab33905) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BCL10 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-bcl10-knockout-hela-cell-lysate-ab257144'>ab257144</a>)

Lane 3:

Ramos cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Predicted band size: 26 kDa

Observed band size: 32 kDa

false

• WB

Lab

Western blot - Anti-Bcl10 antibody [EP606Y] (AB33905)

Lanes 1-4 : Merged signal (red and green). Green - ab33905 observed at 32 kDa. Red - loading control, ab7291 observed at 52 kDa.

ab33905 Anti-Bcl10 antibody [EP606Y] was shown to specifically react with Bcl10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261797 (knockout cell lysate ab257144) was used. Wild-type and Bcl10 knockout samples were subjected to SDS-PAGE. ab33905 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bcl10 antibody [EP606Y] (ab33905) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BCL10 knockout HeLa cell lysate at 20 µg

Lane 3:

Ramos cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Predicted band size: 26 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-Bcl10 antibody [EP606Y] (AB33905)

All lanes:

Western blot - Anti-Bcl10 antibody [EP606Y] (ab33905) at 1/1000 dilution

All lanes:

Recombinant Human Bcl10 protein (<a href='/en-us/products/unavailable/recombinant-human-bcl10-protein-ab82241'>ab82241</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 26 kDa

true

Exposure time: 1min