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Apparent cleavage of poly(ADP-ribose) polymerase in non-apoptotic mouse LTA cells: An artifact of cross-reactive secondary antibody
Authors:Budihardjo  I. Imawati  Poirier   Guy G.  Kaufmann  Scott H.
Affiliation:(1) Division of Oncology Research, Mayo Medical School, Rochester, MN 55905, USA;(2) Department of Pharmacology, Mayo Medical School, Rochester, MN 55905, USA;(3) Poly(ADP-ribose) Metabolism Group, Unit of Health and Environment, CHUL Research Center and Laval University, Québec, G1V 4G2, Canada
Abstract: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 sim89000 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 ~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.
Keywords:poly(ADP-ribose) polymerase  apoptosis  word  antibody cross-reactivity
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