BACKGROUND
RIP (receptor interacting protein) is a 74 kD protein that was detected in a two hybrid screen for proteins capable of interacting with the intracellular domain of the Fas receptor (CD95/APO-1).(1) RIP has an N-terminal kinase domain and a C-terminal death domain that is homologous to the Fas death domain. Over expression of RIP in vitro leads to cell death and like Fas, RIP appears to be an apoptosis-inducing protein. Fas is involved in the apoptotic process of T cell development where it plays an important role in the clonal deletion of self-reactive T cells. In CTL-directed apoptosis, binding of the Fas ligand (Fas L) to Fas results in transduction of a cytolytic signal into the cell that leads to apoptosis. Deletion and mutational studies suggest that the Fas death domain is required for Fas to induce cell death. Interaction of the Fas death domain with other intracellular proteins like RIP are important in the cell death response as downstream components in death signalling pathways.
SPECIFICITY AND PREPARATION
G322-2 recognizes RIP. A truncated recombinant RIP GST fusion protein, lacking the kinase domain of RIP, was used as immunogen. The specificity of the antibody was verified by ELISA, immunoprecipitation and western blot analysis. The antibody is routinely tested by western blot analysis in human Jurkat T cell cells where it recognizes RIP as a 74 kD band. Smaller molecular weight breakdown bands of ~30 kD, 22 kD, and/or 16 kD are also sometimes observed.
USAGE
Applications include immunoprecipitation (1-2 µg/one million cells) and western blot analysis (1-2µg/ml). Jurkat human T cells (ATCC TIB 152) are suggested as a positive control.
REFERENCES
1. Stanger, B.Z., P. Leder, T-H. Lee, E. Kim, and B. Seed. 1995. RIP: A novel protein containing a death domain that interacts with Fas/APO-1 (CD95) in yeast and causes cell death. Cell 81:513-523.
| Cat. No. | Description | Clone | Isotype | Size |
| 65591A | Purified anti-RIP | G322-2 | Mouse IgG1 | 0.1mg |
PharMingen supplies a complete line of antibodies and reagents for apoptosis research, including
Antibodies:
Detection Kits and Reagents
Multi-probe Template sets for RiboQuant RNase Protection Assay
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