BACKGROUND

FADD is a molecule involved in the Fas-mediated cell death pathway (reviewed in 1). Apoptosis is induced when Fas ligand or agonistic Fas antibodies bind to the Fas receptor, and trigger the activation of a cell death signalling pathway. Induction of Fas-mediated apoptosis requires a conserved cytoplasmic motif, referred to as the death domain, that is present in the C-terminal end of Fas. FADD also contains a death domain, and Fas and FADD bind to each other through their respective death domains. Death domains are thought to act as adaptor proteins by linking Fas and other members of the tumor necrosis factor receptor (TNFR) superfamily to downstream signalling pathways. Overexpression of FADD in vitro leads to cell death suggesting that FADD, like Fas, is an apoptosis-inducing protein. The N-terminal, but not the C-terminal death domain, is required for apoptosis induced by FADD overexpression. It is thought that the amino-terminal region of FADD functions by binding to caspase 3 and thereby linking signals from the cell surface to an apoptotic protease cascade.(2) FADD has a calculated molecular weight of 24 kD and migrates at a reduced molecular weight of ~27 kD.

SPECIFICITY AND PREPARATION

A66-2 recognizes human FADD. A recombinant full-length human FADD GST fusion protein was used as immunogen. A66-2 was originally characterized by ELISA, western blot, and immunoprecipitation. It is routinely tested by western blot analysis using Daudi B lymphoma cell lysates.

USAGE

Applications include immunoprecipitation (1-2 µg/one million cells) and western blot analysis (1-2 µg/ml). Daudi B lymphoma cells (ATCC CCL 213) are suggested as a positive control.

REFERENCES
1. Cleveland, J.L. and J.N. Ihle. 1995. Contenders in FasL/TNF death signaling. Cell 81:479-482.
2. Muzio, M., A.M. Chinnaiyan, F.C. Kischkel, et al. 1996. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85:817-827.

Detection Kits and Reagents

• Annexin V FITC
• APO-BRDU™
• APO-DIRECT™

[Homepage] [New Products Page] [Catalog of Products] [Ordering Information] [Technical Service]