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Apparent cleavage of poly(ADP-ribose) polymerase in non-apoptotic mouse LTA cells: An artifact of cross-reactive secondary antibody

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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 ∼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 ~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.

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Authors and Affiliations

  1. Division of Oncology Research, Mayo Medical School, Rochester, MN, 55905, USA

    I. Imawati Budihardjo & Scott H. Kaufmann

  2. Department of Pharmacology, Mayo Medical School, Rochester, MN 55905, USA

    Scott H. Kaufmann

  3. Poly(ADP-ribose) Metabolism Group, Unit of Health and Environment, CHUL Research Center and Laval University, Québec, G1V 4G2, Canada

    Guy G. Poirier

Authors
  1. I. Imawati Budihardjo
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  2. Guy G. Poirier
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  3. Scott H. Kaufmann
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Budihardjo, I.I., Poirier, G.G. & Kaufmann, S.H. Apparent cleavage of poly(ADP-ribose) polymerase in non-apoptotic mouse LTA cells: An artifact of cross-reactive secondary antibody. Mol Cell Biochem 178, 245–249 (1998). https://doi.org/10.1023/A:1006808001462

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