Anti-Bax antibody [EPR18283]

• IP

Supplier Data

Immunoprecipitation - Anti-Bax antibody [EPR18283] (AB182733)

Bax was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182733 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab182733 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate, 10μg (Input).

Lane 2 : ab182733 IP in HeLa whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab182733 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Bax antibody [EPR18283] (ab182733)

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Bax antibody [EPR18283] (AB182733)

Blocking and Diluting buffers : 5% NFDM/TBST

Exposure time 58 s

All lanes:

Western blot - Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell). Whole cell lysates at 20 µg

Lane 2:

Hep G2 (Human hepatocellular carcinoma epithelial cell). Whole cell lysates at 20 µg

Lane 3:

Jurkat (Human T cell leukemia T lymphocyte) Whole cell lysates at 20 µg

Lane 4:

A549 (Human lung carcinoma epithelial cell) Whole cell lysates at 20 µg

Lane 5:

C2C12 (Mouse myoblasts myoblast) Whole cell lysates at 20 µg

Lane 6:

C6 (Rat glial tumor glial cell) Whole cell lysates at 20 µg

Lane 7:

Mouse brain. Whole tissue lysate at 20 µg

Lane 8:

Rat brain. Whole tissue lysate at 20 µg

Lane 9:

Rat Spleen. Whole tissue 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 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 18 kDa

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• WB

Supplier Data

Western blot - Anti-Bax antibody [EPR18283] (AB182733)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1,2,3 and 4 : 8 seconds; Lane 5 and 6 : 1 minute; Lane 7 and 8 : 15 seconds.

All lanes:

Western blot - Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human fetal spleen lysate at 10 µg

Lane 5:

Human fetal lung lysate at 10 µg

Lane 6:

Human thymus lysate at 10 µg

Lane 7:

Human cerebellum lysate at 10 µg

Lane 8:

Human lymph node lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

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• WB

Lab

Western blot - Anti-Bax antibody [EPR18283] (AB182733)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : BAX knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HT29 whole cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab182733 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab182733 was shown to recognize BAX when BAX knockout samples were used, along with additional cross-reactive bands. Wild-type and BAX knockout samples were subjected to SDS-PAGE. ab182733 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bax antibody [EPR18283] (ab182733)

Predicted band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-Bax antibody [EPR18283] (AB182733)

Blocking/Dilution buffer : 5% NFDM/TBST.

Jurkat doesn't express Bax. (PMID : 9583678).

All lanes:

Western blot - Anti-Bax antibody [EPR18283] (ab182733) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

HuT-78 (Human Sezary syndrome cell line) whole cell lysate at 20 µg

Lane 4:

Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 5:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 6:

Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 7:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 8:

L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate at 20 µg

Lane 9:

C2C12 (Mouse myoblast cell line) 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 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Bax antibody [EPR18283] (AB182733)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-7 : 30 seconds; Lane 8-11 : 1 second.

All lanes:

Western blot - Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat kidney lysate at 10 µg

Lane 7:

Rat spleen lysate at 10 µg

Lane 8:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 9:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 10:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 11:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false