Rapid detection of poly(ADP-ribose) polymerase by enzyme-linked immunosorbent assay during its purification and improvement of its purification

We report a new detection method for the purification of poly(ADP-ribose) polymerase (PARP). PARP purification generates many fractions in which PARP is usually detected by a time consuming activity assay. The development of a new method was also needed in order to decrease the utilization of radioa...

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Bibliographic Details
Published in:Molecular and cellular biochemistry 1998-08, Vol.185 (1-2), p.199-203
Main Authors: Sallmann, F R, Plancke, Y D, Poirier, G G
Format: Article
Language:English
Subjects:
Affinity Labels - metabolism
Chromatography, Agarose - methods
Enzyme-Linked Immunosorbent Assay - methods
Heparin - metabolism
Poly(ADP-ribose) Polymerases - isolation & purification
Sepharose - analogs & derivatives
Sepharose - metabolism
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Summary:We report a new detection method for the purification of poly(ADP-ribose) polymerase (PARP). PARP purification generates many fractions in which PARP is usually detected by a time consuming activity assay. The development of a new method was also needed in order to decrease the utilization of radioactivity. This new method, based on an enzyme-linked immunosorbent assay (ELISA), is very rapid, sensitive, and avoids most radioactivity. Moreover, to illustrate this method, a new matrix was used, the Heparin Sepharose. This matrix was chosen for its affinity for the DNA binding proteins and because it allows the separation of whole PARP from its proteolytic fragments.
ISSN:0300-8177
DOI:10.1023/A:1006861015142