Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Apoptosis

DR5 (D4E9) XP® Rabbit mAb #8074

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Endogenous 40, 48 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

DR5 (D4E9) XP® Rabbit mAb recognizes endogenous levels of total DR5 protein. This antibody detects both the short and long isoforms of DR5.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg260 within the cytoplasmic region of human DR5 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using DR5 (D4E9) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from A549 cells, untreated or doxorubicin-treated (500 nM) for the indicated times, using DR5 (D4E9) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with the long isoform of human DR5 (hDR5, +), using DR5 (D4E9) XP® Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

The tumor necrosis factor receptor family, which includes TNF-RI, 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, 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.

DR5 is a receptor for TNF-related apoptosis inducing ligand (TRAIL), which has been shown to induce apoptosis in a variety of cell types and has been targeted for cancer therapy (1-5). Structurally, DR5 contains an amino-terminal leader cleavage site, followed by an extracellular region containing two cysteine-rich repeats, a central transmembrane domain, and a carboxy-terminal DD. DR5 is expressed in a wide variety of tissues and is a transcriptional target of p53 (6-8). It induces apoptosis through a FADD-dependent pathway. Deletion of DR5 leads to resistance in TRAIL-mediated apoptosis as well as an abrogated response to DNA-damaging stimuli (9). At least two isoforms of DR5 are produced by alternative splicing (10).

  1. Nagata, S. (1997) Cell 88, 355-365.
  2. Thorburn, A. (2004) Cell. Signal. 16, 139-144.
  3. Wiley, S.R. et al. (1995) Immunity 3, 673-82.
  4. Walczak, H. et al. (1997) EMBO J 16, 5386-97.
  5. Chaudhary, P.M. et al. (1997) Immunity 7, 821-30.
  6. MacFarlane, M. et al. (1997) J Biol Chem 272, 25417-20.
  7. Wu, G.S. et al. (2000) Adv Exp Med Biol 465, 143-51.
  8. Wu, G.S. et al. (1997) Nat Genet 17, 141-3.
  9. Finnberg, N. et al. (2005) Mol Cell Biol 25, 2000-13.
  10. Screaton, G.R. et al. (1997) Curr Biol 7, 693-6.

Application References

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Companion Products

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

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