Cell Signaling Technology

Product Pathways - Apoptosis

DFF45/DFF35 Antibody #9732

Applications Reactivity Sensitivity MW (kDa) Source
W IP H Endogenous 35, 45 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
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

DFF45/DFF35 Antibody detects endogenous levels of total full length DFF45 and/or full length DFF35. The antibody does not cross-react with related proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp224 of human DFF45. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of Jurkat cells, untreated (control) or etoposide-treated (25 µM), using DFF45/DFF35 Antibody.

Background

Human DFF45 and its mouse homologue ICAD function in normal cells as chaperones for caspase-activated deoxyribonuclease (DFF40 or CAD) during its synthesis (1). The association of DFF45 (or its isoform DFF35) with DFF40 inhibits the DNAse activity of the latter (1-4). In vitro, DFF45 has been shown to be the target of several caspases, including caspase-3, -6, -7, -8 and granzyme B (3). In vivo, caspase-3 is believed to be the primary enzyme responsible for processing DFF45 and release of its carboxy-terminal fragment (3,5). The cleavage of DFF45 inactivates its inhibitory function on DFF40 and causes nuclear DNA degradation by DFF40, leading to cell death (6,7).

  1. Enari, M. et al. (1998) Nature 391, 43-50.
  2. Sakahira, H. et al. (1998) Nature 391, 96-99.
  3. Wolf, B. B. et al. (1999) J. Biol. Chem. 274, 30651-30656.
  4. Gu, J. et al. (1999) J. Biol. Chem. 274, 20759-20762.
  5. Tang, D. and Kidd, V.J. (1998) J. Biol. Chem. 273, 28549-28552.
  6. Liu, X. et al. (1999) J. Biol. Chem. 274, 13836-13840.
  7. Zhang, J. et al. (1999) J. Biol. Chem. 274, 37450-37454.

Application References

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Companion Products

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

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