A general method to express and purify full-length human poly(ADP-ribose) polymerase-1 (PARP-1), individual PARP-1 domains, and groups of PARP-1 domains from Escherichia coli cells is described. The procedure allows for robust production of highly pure PARP-1 that is free of DNA contamination and well-suited for biochemical experiments and for structural and biophysical analysis. Two biochemical assays for monitoring PARP-1 automodification activity are presented that can be used to evaluate purified PARP-1, combinations of PARP-1 domains, or PARP-1 mutants.
CITATION STYLE
Langelier, M. F., Planck, J. L., Servent, K. M., & Pascal, J. M. (2011). Purification of Human PARP-1 and PARP-1 Domains from Escherichia coli for Structural and Biochemical Analysis. In Methods in Molecular Biology (Vol. 780, pp. 209–226). Humana Press Inc. https://doi.org/10.1007/978-1-61779-270-0_13
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