Anti-Cleaved Caspase-7 antibody [EPR22840-25] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-Cleaved Caspase-7 antibody [EPR22840-25] - BSA and Azide free (AB256474)

Cleaved Caspase-7 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1μM Staurosporine for 3h whole cell lysate 10μg with ab256469 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256469. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1μM Staurosporine for 3h whole cell lysate 10μg

Lane 2 : ab256469 IP in HeLa treated with 1μM Staurosporine for 3h whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256469 in HeLa treated with 1μM Staurosporine for 3h whole cell lysate.

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

Exposure time : 15 seconds.

This antibody reacts with an unidentifiable protein around 48 kDa.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256469).

All lanes:

Immunoprecipitation - Anti-Cleaved Caspase-7 antibody [EPR22840-25] (<a href='/en-us/products/primary-antibodies/cleaved-caspase-7-antibody-epr22840-25-ab256469'>ab256469</a>)

Predicted band size: 34 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-Cleaved Caspase-7 antibody [EPR22840-25] - BSA and Azide free (AB256474)

Caspase-7 is activated upon apoptotic induction. Caspase-7 p22 subunit (22 kDa) and p20 subunit (18 kDa) are observed. The molecular weight observed is consistent with what has been described in the literature (PMID : 16352606). This antibody reacts with an unidentifiable protein around 48 kDa. ab256469 was shown to specifically react with Caspase-7 in wild-type HAP1 cells as signal was lost in Caspase-7 knockout cells. Wild-type and Caspase-7 knockout samples were subjected to SDS-PAGE. ab256469 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ instrument using the ECL technique. Blocking and Diluting Buffer and concentration : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256469).

All lanes:

Western blot - Anti-Cleaved Caspase-7 antibody [EPR22840-25] (<a href='/en-us/products/primary-antibodies/cleaved-caspase-7-antibody-epr22840-25-ab256469'>ab256469</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 1 µM staurosporine for 3 hours whole cell lysate at 20 µg

Lane 3:

Untreated wild-type HAP1 whole cell lysate at 20 µg

Lane 4:

Wild-type HAP1 treated with 1 µM staurosporine for 3 hours whole cell lysate at 20 µg

Lane 5:

Untreated caspase-7 knockout HAP1 whole cell lysate at 20 µg

Lane 6:

Caspase-7 knockout HAP1 treated with 1 µM staurosporine for 3 hours whole cell lysate at 20 µg

Lane 7:

Mouse spleen lysate at 20 µg

Lane 8:

Rat spleen 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: 34 kDa

Observed band size: 18 kDa,22 kDa

false

Exposure time: 26s