Anti-Caspase-9 antibody [E84] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] - BSA and Azide free (AB247220)

This data was developed using ab32068, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-Pro-Caspase-9 RabMAb (ab32068).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-Caspase-9 antibody [E84] - BSA and Azide free (AB247220)

This data was developed using ab32068, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Caspase-9 antibody [E84] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-e84-ab32068'>ab32068</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with staurosporine 1uM for 4 hours 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 1/20000 dilution

Predicted band size: 46 kDa

Observed band size: 35 kDa,37 kDa,46 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Caspase-9 antibody [E84] - BSA and Azide free (AB247220)

This data was developed using ab32068, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 whole cell lysate (40 μg).

Lane 2 : CASP9 knockout HAP1 whole cell lysate (40μg).

Lane 3 : HeLa whole cell lysate (40 μg).

Lanes 1 - 3 : Merged signal (red and green). Green - ab32068 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32068 was shown to specifically recognize CASP9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CASP9 knockout samples were examined. Wild-type and CASP9 knockout samples were subjected to SDS-PAGE. ab32068 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caspase-9 antibody [E84] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-e84-ab32068'>ab32068</a>)

Predicted band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-Caspase-9 antibody [E84] - BSA and Azide free (AB247220)

This data was developed using ab32068, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Caspase-9 antibody [E84] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-e84-ab32068'>ab32068</a>) at 1/2000 dilution

All lanes:

Jurkat cell lysate at 10 µg

Predicted band size: 46 kDa

Observed band size: 46 kDa

false