Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information
This antibody only detects the active (cleaved) form of Caspase-3 and does not recognize the pro form of Caspase-3.
PubMedID 19789217 describes the detection of human cells injected into mice.
IHC-Fr, WB, ICC/IFmore details Unsuitable for:
Flow Cyt or IP
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human active Caspase-3 aa 1-100 (N terminal). A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit). Database link: P42574
WB: Jurkat cell lysate (camptothecin treated); HeLa cell lysate (staurosporine treated).
IF: Hela cells (staurosporine treated).
This product is a recombinant rabbit monoclonal antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).
1/100 - 1/250.
Is unsuitable for Flow Cyt or IP.
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp- -Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.
Belongs to the peptidase C14A family.
Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa. S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.
Western blot - Anti-active Caspase-3 antibody [E83-77] (ab32042)
Predicted band size : 32 kDa
Lane 1: Wild type HAP1 + DMSO for 24 hours, whole cell lysate (20 µg) Lane 2: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg) Lane 3: HAP1 CASP3 KO + DMSO for 24 hours, whole cell lysate (20 µg) Lane 4: HAP1 CASP3 KO + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg) Lane 5: HeLa + DMSO for 24 hours, whole cell lysate (20 µg) Lane 6: HeLa + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg) Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, ab8245, observed at 130 kDa.
ab32042 was shown to specifically react with CASP3 (Caspase-3) when CASP3 (Caspase-3) knockout samples were used. HAP1 wild-type and CASP3 (Caspase-3) knockout samples were subjected to SDS-PAGE. Ab32042 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 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 1/10000 dilution for 1 hour at room temperature before imaging.
Cells were grown to confluency prior to treatment.