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Poly(ADP-ribose) Preparation Using Anion-Exchange Column Chromatography

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Abstract

Poly(ADP-ribose) polymerase (PARP) polyADP-ribosylates proteins involved in various physiological processes. Accumulated evidence suggests not only protein-conjugated poly(ADP-ribose) but also protein-free poly(ADP-ribose) function in various physiological processes. There are increasing occasions that require protein-free poly(ADP-ribose) to study the function and dynamics of poly(ADP-ribose) in cells. However, the availability of poly(ADP-ribose) is still limited because a chemical synthesis method has not been established. Here, we describe an improved method for the preparation of protein-free poly(ADP-ribose), synthesized enzymatically by using a recombinant PARP-1 expression system and purified with an anion-exchange column chromatography. This method will be useful for biochemical and biological investigation of poly(ADP-ribose) functions and dynamics.

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Key concepts: Poly ADP ribose polymerase, Ribose, Ion chromatography, Biochemistry, Polymerase, Ion exchange, Chemistry, Function (biology)