Caspase 3 is one of the key executioners of apoptosis. It is responsible either partially or totally for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP). Activation of caspase 3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 subunits. Cleavage of caspase 3 requires aspartic acid at the P1 position.   Suitable for use in Flow Cytometry, Western Blot, Immunoprecipitation, Immunohistochemistry and Immunofluorescence/Immunocytochemistry. Other applications not tested.

Flow Cytometry: 1:800 Western Blot: 1:1000 Incubate membrane with diluted antibody in 5% nonfat dry milk, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Immunohistochemistry (Paraffin): 1:400 Requires HIER using sodium citrate buffer pH6.0 prior to staining. Immunohistochemistry (Frozen): 1:400 fixed in 3% Formaldehyde. Immunofluorescence (IC): 1:400 Immunoprecipitation: 1:100 Optimal dilutions to be determined by the researcher.

May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Synthetic peptide corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3. |Species Sequence Homology: porcine, bovine and canine

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Purified by Protein A and immunoaffinity chromatography.

Recognizes cleaved human Caspase 3 (Asp175). Detects endogenous levels of the large fragment (17/19kD) of activated caspase-3 resulting from cleavage adjacent to Asp175. Does not recognize full length caspase-3 or other cleaved caspases. Detects non-specific caspase sunstrates by Western Blot.|Species Crossreactivity: mouse, rat and monkey.