Cell Signaling Technology

Product Pathways - Apoptosis

Apaf-1 (D5C3) Rabbit mAb #8969

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M (Mk) Endogenous 135 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Apaf-1 (D5C3) Rabbit mAb recognizes endogenous levels of total Apaf-1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro470 of human Apaf-1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Apaf-1 siRNA I #6229 (+), using Apaf-1 (D5C3) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Apaf-1 (D5C3) Rabbit mAb confirms silencing of Apaf-1 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Apaf-1 (D5C3) Rabbit mAb.

Background

Apoptotic protease activating factor 1 (Apaf-1), originally identified as the mammalian homolog of the C. elegans apoptotic regulatory protein CED-4, is an important signaling protein involved in the activation of caspase-9 during apoptosis (1). Cytosolic Apaf-1 forms a complex with caspase-9 in the presence of cytochrome c and dATP, ultimately leading to caspase-9 activation and subsequent activation of caspase-3 (2,3). The protein contains an amino-terminal CARD domain, a central CED-4 homology domain, and multiple WD-40 repeats at the carboxy-terminus. Several isoforms of Apaf-1 are expressed through alternative splicing generating a small insert following the CARD domain as well as an extra WD-40 repeat (4). Apaf-1 knock-out mice display widespread defects in apoptosis and resistance to a variety of apoptotic stimuli (5,6).

  1. Zou, H. et al. (1997) Cell 90, 405-13.
  2. Zou, H. et al. (1999) J Biol Chem 274, 11549-56.
  3. Saleh, A. et al. (1999) J Biol Chem 274, 17941-5.
  4. Benedict, M.A. et al. (2000) J Biol Chem 275, 8461-8.
  5. Cecconi, F. et al. (1998) Cell 94, 727-37.
  6. Yoshida, H. et al. (1998) Cell 94, 739-50.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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