Anti-Caspase-9 antibody [EPR18868]

• IP

Supplier Data

Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (AB184786)

Caspase 9 was immunoprecipitated from 1mg of NIH/3T3 whole cell lysate (Mouse embyro fibroblast cells) treated with 1μM staurosporine for 4h with ab184786 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184786 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h,10ug (Input).
Lane 2 : ab184786 IP in NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab184786 in NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 10 seconds.

The product has better affinity to procaspase-9. The cleaved form p37 could not be observed even with 3 minutes exposure time.

All lanes:

Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)

Predicted band size: 46 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (AB184786)

Caspase 9 was immunoprecipitated from 1mg of C6 whole cell lysate (Rat glial tumor cells) treated with 1μM staurosporine for 4h with ab184786 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184786 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : C6 whole cell lysate treated with 1μM staurosporine for 4h, 10ug (Input).
Lane 2 : ab184786 IP in C6 whole cell lysate treated with 1μM staurosporine for 4h.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab184786 in C6 whole cell lysate treated with 1μM staurosporine for 4h.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

The product has better affinity to procaspase-9. The cleaved form p37 could be observed with longer exposure time.

All lanes:

Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)

Predicted band size: 46 kDa

false

• WB

Supplier Data

Western blot - Anti-Caspase-9 antibody [EPR18868] (AB184786)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (Mouse embyro fibroblast cell line) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 whole cell lysate treated with 1 µM staurosporine for 4 hours 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: 46 kDa

Observed band size: 37 kDa,39 kDa,50 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Caspase-9 antibody [EPR18868] (AB184786)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat heart 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: 46 kDa

Observed band size: 37 kDa,39 kDa,50 kDa

false

Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-Caspase-9 antibody [EPR18868] (AB184786)

Blocking/Dilution buffer : 5% NFDM/TBST.

The p39 and p37 subunits were inhibited when the caspase activity was blocked by the caspase inhibitor, Z-VAD-FMK.

All lanes:

Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

Lane 1:

Untreated C6 (Rat glial tumor cells) whole cell lysate at 20 µg

Lane 2:

C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 0.5 hour at 20 µg

Lane 3:

C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 1 hour at 20 µg

Lane 4:

C6 whole cell lysate treated with 1 µm staurosporine for 4 hours at 20 µg

Lane 5:

C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 1 hour, then treated with 1 µM staurosporine for 4 hours 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: 46 kDa

Observed band size: 37 kDa,39 kDa,50 kDa

false

Exposure time: 10s