Anti-Caspase-3 抗体

• WB

Lab

Western blot - Anti-Caspase-3 antibody (AB13847)

Caspase 3 exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce large (17kDa) and small (12kDa) subunits. These subunits dimerize to form the active enzyme. ab13847 specifically detects the large active subunit (17kDa) and the inactive pro Caspase 3 (32 kDa).

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab13847 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

Secondary antibody - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody

All lanes:

Western blot - Anti-Caspase-3 antibody (ab13847) at 1 µg/mL

Lane 1:

Human Caspase 3 (active) Recombinant Protein at 0.1 µg

Lane 2:

Human Pro Caspase 3 (inactive) Recombinant Protein at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 31 kDa

Observed band size: 17 kDa,32 kDa

true

Exposure time: 8min

• WB

Lab

Western blot - Anti-Caspase-3 antibody (AB13847)

Lane 1 : Wild-type HAP1 cell lysate + Staurosporine (ab146588) (1μM for 4h)
Lane 2 : Wild-type HAP1 cell lysate
Lane 3 : Caspase-3 knockout HAP1 cell lysate + Staurosporine (ab146588) (1μM for 4h)
Lane 4 : Caspase-3 knockout HAP1 cell lysate

Lanes 1 - 4 : Merged signal (red and green). Green - ab13847 observed at 17 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab13847 was shown to recognise Caspase 3 when Caspase 3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase 3 knockout samples (±staurosporine treatment) were subjected to SDS-PAGE. ab13847 and ab8245 (loading control to GAPDH) were diluted to 1/500 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caspase-3 antibody (ab13847)

Predicted band size: 31 kDa

Observed band size: 26 kDa,34 kDa

false

• WB

AbReview72311****

Western blot - Anti-Caspase-3 antibody (AB13847)

4-20% (Mini protean gels) non-denaturing conditions

All lanes:

Western blot - Anti-Caspase-3 antibody (ab13847) at 1/1000 dilution

Lane 1:

Skin (atopic dermatits) whole cell lysate at 20 µg

Lane 2:

Skin (without atopic dermatits) whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti rabbit HRP at 1/2000 dilution

false

Exposure time: 5min

This image is courtesy of an anonymous Abreview