The Apoptosis Antibody Sampler Kit (Mouse Specific) is designed for use with mouse samples and offers an economical way of testing for apoptosis. The kit contains 40ul of each primary antibody and 100ul of Anti-rabbit IgG, HRP-linked Antibody. Apoptosis, or programmed cell death, is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10 and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, alpha-Fodrin, DFF and Lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase. Proapoptotic stimuli include the FasL, TNF-alpha, DNA damage and ER stress. Fas and TNFR activate caspases 8 and 10; DNA damage leads to the activation of caspase-9; and ER stress leads to the calcium-mediated activation of caspase-12. The inhibitor of apoptosis protein (IAP) family includes XIAP and Survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of the IAPs on caspases (6).
1 Kit (8 x 40ul)
1. IgG(HRP)Gt x Rb 1x100ul
2. Caspase-3 Pab Rb x 1x40ul
3. Cleaved Caspase-3 (Asp175) Pab Rb x 1x40ul
4. Cleaved Caspase-6 (Asp162) Pab Rb x 1x40ul
5. Caspase-6 Pab Rb x 1x40ul
6. Cleaved PARP (Asp214) RbxMo 1x40ul
7. Caspase-12 Pab Rb x 1x40ul
8. Caspase-9 Pab Rb x Mo 1x40ul
9. Cleaved Caspase-9 (Asp353) Pab Rb x Mo 1x40ul
Each antibody in the kit detects endogenous levels of its respective protein. Cleaved Caspase-3 (Asp175) antibody and Cleaved Caspase-6 (Asp162) antibody detect the large fragment of their respective proteins. Caspase-3 antibody and Caspase-9 antibody (Mouse Specific) detect the full length and the large fragments. Cleaved caspase-9 (Asp353) detects the cleaved large 37 kD fragment. Caspase-6 antibody detects the full length fragment and the small subunit. Caspase-12 antibody detects the full length fragment and its cleaved product. Cleaved PARP (Asp214) antibody (Mouse Specific) detects the large fragment of mouse PARP produced by caspase cleavage.
Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH coupled) corresponding to amino-terminal residues of proteolytic cleavage sites of cleaved caspase-3 (Asp175), cleaved caspase-6 (rat Asp162), cleaved caspase-9 (Asp353), mouse-cleaved PARP, and residues surrounding amino acid 158 of mouse caspase-12. Antibodies are purified by protein A and peptide affinity chromatography.