The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
Is unsuitable for Flow Cyt or IP.
Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp- -Gly-217' bond. Overexpression promotes programmed cell death.
Highly expressed in lung, skeletal muscle, liver, kidney, spleen and heart, and moderately in testis. No expression in the brain.
Belongs to the peptidase C14A family.
Cleavages by granzyme B or caspase-10 generate the two active subunits. Propeptide domains can also be cleaved efficiently by caspase-3. Active heterodimers between the small subunit of caspase-7 and the large subunit of caspase-3, and vice versa, also occur.
Western blot - Anti-pro Caspase 7 antibody [Y33] (ab32067)
Predicted band size: 34 kDa
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: Staurosporin treated HAP1 whole cell lysate (20 µg) Lane 3: CASP7 knockout HAP1 whole cell lysate (20 µg) Lane 4: Staurosporin treated CASP7 knockout HAP1 whole cell lysate (20 µg) Lane 5: HeLa whole cell lysate (20 µg) Lane 6: Staurosporin treated HeLa whole cell lysate (20 µg)
Lanes 1 - 6: Merged signal (red and green). Green - ab32067 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab32067 was shown to recognize pro Caspase 7 when HAP1 + pro Caspase 7 knockout samples were used, along with additional cross-reactive bands. Wild-type and pro Caspase 7 knockout samples were subjected to SDS-PAGE. Ab32067 and ab18058 (Mouse anti-Vinculin 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 1/20000 dilution for 1 hour at room temperature before imaging.