Death Receptor Antibody Sampler Kit II #29603

Kit Usage Information

Protocols

• 3727: Western Blotting, Immunoprecipitation (Magnetic)
• 3736: Western Blotting, Immunoprecipitation (Magnetic)
• 4233: Western Blotting, Immunohistochemistry (Paraffin)
• 7074: Western Blotting
• 8074: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence
• 20772: Western Blotting
• 42533: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow
• 93026: Western Blotting, Immunoprecipitation (Agarose)

Product Description

The Death Receptor Antibody Sampler Kit II provides an economical means to investigate members of the death receptor family. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Death Receptor Antibody Sampler Kit II detects endogenous levels of its target protein. TNF-R1 (C25C1) Rabbit mAb recognizes a 30 kDa splice isoform of TNF-R1 in some cell lines. DR3 (D4O3X) Rabbit mAb detects a band unknown origin at 110 kDa in some cell lines.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Lys259 of human Fas, Ser331 of human TNF-R1, Leu335 of human DR3, Arg260 of human DR5, His235 of human DR6, or a recombinant protein within the carboxy terminus of DR4. The polyclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp339 of human TNF-R2. Antibodies are purified by protein A and peptide affinity chromatography.

Background

The tumor necrosis factor receptor family, which includes TNF-RI, TNF-R2, Fas, DR3, DR4, DR5, and DR6, plays an important role in the regulation of apoptosis in various physiological systems (1,2). The receptors are activated by a family of cytokines that include TNF, FasL, TWEAK, and TRAIL. They are characterized by a highly conserved extracellular region containing cysteine-rich repeats and a conserved intracellular region of about 80 amino acids termed the death domain (DD). The DD is important for transducing the death signal by recruiting other DD containing adaptor proteins (FADD, TRADD, RIP) to the death-inducing signaling complex (DISC) resulting in activation of caspases. The two receptors for TNF-α, TNF-R1 (55 kDa) and TNF-R2 (75 kDa) can mediate distinct cellular responses (3,4). In most cases cytotoxicity elicited by TNF has been reported to act through TNF-R1 (5,6). DR3/WSL-1/Apo-3/TRAMP/LARD is a TNFR family member containing the characteristic extracellular cysteine-repeats, transmembrane region, and an intracellular DD (7-11). DR3 is activated by its ligand Apo-3L/TWEAK to induce apoptosis and activation of NF-κB (12,13). Like TNF-R1, DR3 binds to the DD adaptor protein TRADD, which can then associate with other DD proteins like FADD and RIP as well as members of the TRAF family (7,8). Tissue expression of DR3 is very restricted, primarily seen on the surface of activated thymocytes and lymphocytes and plays an important role in thymocyte negative selection (7,8,14). Studies have also indicated an association with DR3 and rheumatoid arthritis (15,16). DR4 (TRAIL-RI, TNFRSF10A) and DR5 (TRAIL-R2, TNFRSF10B) are receptors for the cytokine TRAIL. Both receptors contain death domains that recruit DISC complexes triggering caspase activation and apoptosis (17-20). DR6, also known as TNFRSF21, is a TNFR family member able to induce apoptosis as well as activation of NF-κB and JNK (21). DR6 appears to play a critical role in the activation and differentiation of T and B lymphocytes (22,23). In the nervous system, β-amyloid precursor protein (APP) activates DR6 to trigger neuronal degeneration (24).

1. Nagata, S. (1997) Cell 88, 355-65.
2. Thorburn, A. (2004) Cell Signal 16, 139-44.
3. Tartaglia, L.A. et al. (1991) Proc Natl Acad Sci U S A 88, 9292-6.
4. Peschon, J.J. et al. (1998) J Immunol 160, 943-52.
5. Tartaglia, L.A. et al. (1993) Cell 73, 213-6.
6. Rothe, J. et al. (1993) Nature 364, 798-802.
7. Chinnaiyan, A.M. et al. (1996) Science 274, 990-2.
8. Kitson, J. et al. (1996) Nature 384, 372-5.
9. Marsters, S.A. et al. (1996) Curr Biol 6, 1669-76.
10. Bodmer, J.L. et al. (1997) Immunity 6, 79-88.
11. Screaton, G.R. et al. (1997) Proc Natl Acad Sci U S A 94, 4615-9.
12. Marsters, S.A. et al. (1998) Curr Biol 8, 525-8.
13. Kaptein, A. et al. (2000) FEBS Lett 485, 135-41.
14. Wang, E.C. et al. (2001) Mol Cell Biol 21, 3451-61.
15. Osawa, K. et al. (2004) Genes Immun 5, 439-43.
16. Borysenko, C.W. et al. (2005) Biochem Biophys Res Commun 328, 794-9.
17. Pan, G. et al. (1997) Science 276, 111-3.
18. Walczak, H. et al. (1997) EMBO J 16, 5386-97.
19. Chaudhary, P.M. et al. (1997) Immunity 7, 821-30.
20. Schneider, P. et al. (1997) Immunity 7, 831-6.
21. Pan, G. et al. (1998) FEBS Lett 431, 351-6.
22. Zhao, H. et al. (2001) J Exp Med 194, 1441-8.
23. Schmidt, C.S. et al. (2003) J Exp Med 197, 51-62.
24. Nikolaev, A. et al. (2009) Nature 457, 981-9.

Pathways & Proteins

Explore pathways + proteins related to this product.

Database Links

UniProt ID: Q93038 , O75509 , P25445 , P19438 , P20333 , O00220 , O14763

Entrez-Gene Id: 8718 , 27242 , 355 , 7132 , 7133 , 8797 , 8795

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