Product Pathways - Apoptosis
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625
PhosphoSitePlus® protein, site, and accession data: PARP1
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H Mk | Endogenous | 89 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
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 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.
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with TNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (upper), or total PARP Antibody #9542 (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (green). Actin filament were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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).
- Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
- Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
- Cohen, G.M. (1997) Biochem. J. 326, 1-16.
- Nicholson, D. W. et al. (1995) Nature 376, 37-43.
- Tewari, M. et al. (1995) Cell 81, 801-809.
- Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.
Application References
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Companion Products
- 9541 Cleaved PARP (Asp214) Antibody (Human Specific)
- 9542 PARP Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 8112 SignalStain® Antibody Diluent
- 5425 Normal Goat Serum
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
- 8104 SignalSlide® Cleaved Caspase-3 (Asp175) IHC Controls
For Research Use Only. Not For Use In Diagnostic Procedures.
