Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Apoptosis

Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #6987

Applications Reactivity Sensitivity Isotype
F H Mk Endogenous Rabbit IgG

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of the large fragment (89 kDa) of human PARP1 protein produced by caspase cleavage. The antibody does not recognize full length PARP1 or other PARP isoforms.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp214 in human PARP protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved PARP (Asp214) (D64E10) XP® (Alexa Fluor® 647 Conjugate) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with staurosporine #9953 (1 μM for 3 hours, right), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3623 (green).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625.

Background

PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (1). This protein can be cleaved by many ICE-like caspases in vitro (2,3) and is one of the main cleavage targets of caspase-3 in vivo (4,5). In human PARP, the cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (2,4). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (6).

  1. Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
  2. Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
  3. Cohen, G.M. (1997) Biochem. J. 326, 1-16.
  4. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
  5. Tewari, M. et al. (1995) Cell 81, 801-809.
  6. Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.

Application References

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

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