Polyclonal Antibody to Apaf 1

Background
Principal Names: Apoptotic protease activating factor 1 (APAF1); Apaf-1; CED4; KIAA0413; Official Gene Symbol: APAF1 Gene ID: 317 (Human) Gene Map Locus: 12q23 (Human)

Apaf-1, a 135KDa member of NOD family proteins, is a mammalian homolog of C.elegans CED4. It consists of an N-terminal caspase recruitment domain (CARD), a central nucleotide-binding oligomerization domain (NOD) and multiple WD40 repeats at the C-terminus. Apaf-1 exists as an inactive protein in the cells and requires ATP for activation. It binds with cytochrome C and dATP and oligomerizes into a large apoptosome complex responsible for the activation of procaspase-9. Activated caspase 9 stimulates the subsequent caspase cascade that commits the cell to apoptosis. Apaf-1 is an essential downstream effector of p53-mediated apoptosis and is inactivated in metastatic melanomas.

Antigen
A synthetic peptide derived from C-terminus of human Apaf 1.

Application Notes
We suggest an incubation period of 30 minutes at room temperature. However, depending upon the fixation conditions and the staining system employed, optimal incubation conditions should be determined by the user. Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining. Specificity: Apaf-1 is a cytosolic protein that rests in a latent state until bound by cytochrome c (Apaf-2). The resulting Apaf-1/cytochrome c complex associates with the zymogen form of caspase-9 (Apaf-3) in the presence of dATP or ATP, promoting the autocatalytic activation of caspase-9. This antibody will be useful in studies of the activation of caspases through Apaf-1 and also in metastatic melanomas where Apaf-1 is inactivated which leads to defects in the execution of apoptotic cell death. This antibody cross reacts with mouse and rat. Cellular Localization: Cytoplasmic.

Genebank Info (Protein)
NP_863659

Genebank Info (Nucleotide)
NP_863651

Reference
1. Reidl et.al Nature 434; 926-933 (14 April, 2005)
2. Chu et.al JBC 276 (12) 9239-9245 (23 March, 2001)
3. Robles et.al Cancer Res 61; 6660-6664 (2001)