Anti-Bax antibody [EPR18284]

• WB

Lab

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Lanes 1 - 4 : Merged signal (red and green). Green - ab182734 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab182734 was shown to specifically react with BAX when BAX knockout samples were used. Wild-type and BAX knockout samples were subjected to SDS-PAGE. ab182734 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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 [EPR18284] (ab182734) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

BAX knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

HT29 whole cell lysate at 20 µg

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Blocking and diluting 5% NFDM/TBST

Exposure time 1~9 lanes 24 s; 10~11 lanes 3 min

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734) 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

Lanes 8 and 10:

Rat brain. Whole tissue lysate at 20 µg

Lanes 9 and 11:

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 [EPR18284] (AB182734)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1, 2 and 3 : 3 seconds; Lane 4 : 2 seconds; Lane 5 : 3 seconds; Lane 6 and 7 : 2 seconds; Lane 8, 9 and 10 : 1 second.

This product reacts with a 15 kD isoform (PMID : 7607685).

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 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:

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

Lane 10:

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: 19 kDa

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• IP

Supplier Data

Immunoprecipitation - Anti-Bax antibody [EPR18284] (AB182734)

Bax was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182734 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab182734 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 : ab182734 IP in HeLa whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab182734 in HeLa whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Bax antibody [EPR18284] (ab182734)

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Lanes 1- 2 : Merged signal (red and green). Green - ab182734 observed at 21 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab182734 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BAX knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAX knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-bax-knockout-hela-cell-line-ab255363'>ab255363</a>)

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Lanes 1 - 2 : Merged signal (red and green). Green - ab182734 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa or ab18058, observed at 130 kDa.

This western blot image is a comparison between ab182734 and a competitor's top cited rabbit polyclonal antibody.

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734)

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

Bax knockout HAP1 cell lysate at 20 µg

Predicted band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Blocking/Dilution buffer : 5% NFDM/TBST.

This product reacts with a 15 kD isoform (PMID : 7607685).

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 dilution

Lane 1:

Human kidney lysate at 10 µg

Lane 2:

Human spleen 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: 19 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Blocking/Dilution buffer : 5% NFDM/TBST.

Negative control :

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

All lanes:

Western blot - Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 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: 19 kDa

false

Exposure time: 2s

• WB

CiteAb

Western blot - Anti-Bax antibody [EPR18284] (AB182734)

Bax western blot using anti-Bax antibody [EPR18284] ab182734. Publication image and figure legend from Zhang, X. Z., Zhang, M. H., et al., 2019, Biosci Rep, PubMed 31769476.

ab182734 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab182734 please see the product overview.

Expression of Bcl-2, BAX and Caspase-3 in rat gastric smooth muscle cells under high glucose and AMPK silencing conditions(A) Expression of Bcl-2 in rat gastric smooth muscle cells under high glucose and AMPK silencing conditions. ***p<0.001, vs high glucose-24h group, **p<0.001, vs the high glucose-48h group. (B) Expression of BAX in rat gastric smooth muscle cells under high glucose and AMPK silencing conditions. *p<0.05, vs high glucose-24h group. (C) Expression of Caspase-3 in rat gastric smooth muscle cells under high glucose and AMPK silencing conditions. *p<0.05, vs high glucose-24h group.

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