Technical Data
APAF1, CT (Apoptotic Protease-activating Factor 1, APAF-1, KIAA0413)
Apaf 1 (apoptosis protease-activating factor-1) is a key regulator of the mitochondrial apoptotic pathway, being the central element of the multimeric apoptosome. The apoptosome consists of cytochrome c, procaspase-9 and seven Apaf 1 monomers, and is considered to be core apoptotic machinery that executes mitochondria-dependent apoptosis. Cytochrome c, normally compartmentalized in the mitochondria, is released into the cytoplasm following apoptotic stimuli. Apaf 1 binds cytochrome c in the cytoplasm and in the presence of DTP/ATP forms the apoptosome. The apoptosome binds procaspase-9 and promotes its autocatalytic activation. Active caspase-9, in turn, activates downstream caspases including 3, 6, and 7 contributing to the proteolytic caspase activation cascade which leads to cell death. Apaf 1 is a ~130kD protein. Alternative Apaf 1 splicing isoforms of Apaf 1 have been identified in mammalian cells, each having either or both an 11aa insertion in the N-terminus and an additional WD40 repeat (43aa) in the C-terminus. Additionally, Apaf 1 can be proteolytically processed by active caspase-3 during apoptosis resulting in an Apaf 1 cleavage fragment of ~84kD.

Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-1ug/ml
Immunohistochemistry (paraffin): 20ug/ml
Optimal dilutions to be determined by the researcher.

Positive Control:
HeLa or human heart tissue lysate

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
PabIgGAffinity Purified
50ug-20CBlue IceHumanRabbit
As reported
Synthetic peptide corresponding to aa1158-1177 of human Apaf 1, TFYTNGTNLKKIHVSPDFKT.
Purified by Protein G affinity chromatography.
Supplied as a liquid in PBS, 0.02% sodium azide.
Recognizes human Apaf 1 (CT). Species Crossreactivity: mouse and rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Zou H, Henzel WJ, Liu X, Lutschg A, Wang X. Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 1997;90:405-13 2. Cecconi F, Alvarez-Bolado G, Meyer BI, Roth KA, Gruss P. Apaf1 (CED-4 homolog) regulates programmed cell death in mammalian development. Cell 1998;94:727-37 3. Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997;91:479-89 4. Hu Y, Benedict MA, Wu D, Inohara N, Nunez G. Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation. Proc Natl Acad Sci.