Apparent cleavage of poly(ADP-ribose) polymerase in non-apoptotic mouse LTA cells: an artifact of cross-reactive secondary antibody.
Proteolytic cleavage of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) to fragments of 89 kD and 24 kD is widely observed during apoptotic cell death. In the present study, labelling of a Mr approximately 89000 polypeptide was demonstrated in untreated mouse LTA cells during probing of immunoblots with C-2-10 monoclonal anti-PARP antibody. The source of the labeling was traced to the secondary antibody preparation, which labeled a Mr approximately 89000 polypeptide in murine LTA cells but not in human cells. These observations indicate that assessment of PARP cleavage must be (1) performed with appropriate controls when new cell lines are investigated and (2) carefully interpreted in light of additional biochemical or morphological data demonstrating apoptotic changes.
Budihardjo
I I
II
Division of Oncology Research, Mayo Clinic, and Mayo Medical School, Rochester, MN 55905, USA.
Poirier
G G
GG
Kaufmann
S H
SH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017209:Apoptosis; D006690:Bisbenzimidazole / Q000378:metabolism; D002460:Cell Line; D003429:Cross Reactions; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology* / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D011065:Poly(ADP-ribose) Polymerases; D006690:Bisbenzimidazole