Technical Data
A2298-65C
APAF1 (Apoptotic Protease-activating Factor 1, APAF-1, KIAA0413)
Description:
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.

Applications:
Suitable for use in Western Blot, Immunohistochemistry and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:2000
Immunohistochemistry (formalin fixed paraffin embedded): 1:1000-1:5000
Immunoprecipitation: 1:50-1:200
Immunohistochemistry: Frozen
Optimal dilutions to be determined by the researcher.

Positive Control:
Spleen, lymphoid, many cancer cell lines

Storage and Stability:
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 at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGSerum
SizeStorageShippingSourceHost
50ul-20°CBlue IceHumanRabbit
Concentration:
Not determined.
Immunogen:
Synthetic peptide corresponding to aa264-282 (GPKYVVPVESSLGKEKGLE) of human Apaf 1; GenBank no. NP_001151.1.
Purity:
Serum
Form
Supplied as a liquid, 0.05% sodium azide.
Specificity:
Recognizes human Apaf 1.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Campioni M, D Stantini, G Tonini, R Murace, E Dragonetti, EP Spugnini, A Baldi. 2005. Role of Apaf-1, a key regulator of apoptosis, in melnoma progression and chemoresistance. Experimental Dermatology 14:811-818. 2. Lauber K, AE HAE Appel, SF Schlosser, M Gregor, K Schulze-Osthoff, and S Wesselborg. 2001. The adapter protein apoptotic protease-activating factor-1 (Apaf-1) is proteolytically processed during apoptosis. J Biol Chem. 276:29772-29781.