PMID- 11314271
OWN - NLM
STAT- MEDLINE
DCOM- 20010823
LR  - 20180928
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 30
IP  - 4
DP  - 2001 Apr
TI  - Poly (ADP-ribose) polymerase cleavage monitored in situ in apoptotic cells.
PG  - 886-91
AB  - During apoptosis, the activation of a family of cysteine proteases, or caspases, 
      results in proteolytic cleavage of numerous substrates. Antibody probes specific 
      for neoepitopes on protein fragments generated by caspase cleavage provide a 
      means to monitor caspase activity at the level of the individual cell. Poly 
      (ADP-ribose) polymerase (PARP), a nuclear enzyme involved in DNA repair, is a 
      well-known substrate for caspase-3 cleavage during apoptosis. Its cleavage is 
      considered to be a hallmark of apoptosis. Here, we demonstrate that an 
      affinity-purified polyclonal antibody to the p85 fragment of PARP is specific for 
      apoptotic cells. Western blots show that the antibody recognizes the 85-kDa (p85) 
      fragment of PARP but not full-length PARP. We demonstrate a time course of PARP 
      cleavage and DNA fragmentation in situ using the PARP p85 fragment antibody and 
      terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in 
      Jurkat cells treated with anti-Fas. Furthermore, our results indicate that the 
      p85 fragment of PARP resulting from caspase cleavage during apoptosis is rapidly 
      localized outside the condensed chromatin but not in the cytoplasm.
FAU - O'Brien, M A
AU  - O'Brien MA
AD  - Promega Corporation, Madison, WI, USA. mobrien@promega.com
FAU - Moravec, R A
AU  - Moravec RA
FAU - Riss, T L
AU  - Riss TL
LA  - eng
PT  - Journal Article
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (Antibodies)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Indoles)
RN  - 0 (fas Receptor)
RN  - 47165-04-8 (DAPI)
RN  - EC 2.4.2.30 (Poly(ADP-ribose) Polymerases)
SB  - IM
MH  - Antibodies
MH  - Antibody Specificity
MH  - *Apoptosis
MH  - Blotting, Western
MH  - Cytoplasm/chemistry
MH  - DNA Fragmentation
MH  - Fluorescent Dyes
MH  - Humans
MH  - *In Situ Nick-End Labeling
MH  - Indoles
MH  - Jurkat Cells
MH  - Poly(ADP-ribose) Polymerases/immunology/*metabolism
MH  - fas Receptor/analysis/immunology
EDAT- 2001/04/21 10:00
MHDA- 2001/08/24 10:01
CRDT- 2001/04/21 10:00
PHST- 2001/04/21 10:00 [pubmed]
PHST- 2001/08/24 10:01 [medline]
PHST- 2001/04/21 10:00 [entrez]
AID - 10.2144/01304pf01 [doi]
PST - ppublish
SO  - Biotechniques. 2001 Apr;30(4):886-91. doi: 10.2144/01304pf01.