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Pull down of purified, bacterially expressed PP1 isoforms via His-tagged RNA Helicase Ddx21.

By Veerle De Wever (155261), David C. Lloyd (155265), Isha Nasa (155269), Mhairi Nimick (155273), Laura Trinkle-Mulcahy (13747), Robert Gourlay (155277), Nick Morrice (79136) and Greg B. G. Moorhead (155283)

Abstract

<p>Bacterially expressed and purified PP1 (α, β, γ) and His6-Ddx21 (wt, motif1, motif2, double) were incubated together and then mixed with precleared Ni-NTA beads, all at 4°C. Each PP1 isoform was also incubated without Ddx21 (-) to verify unspecific binding of PP1 to the Ni-NTA beads. Enriched His6-Ddx21 and co-precipitated PP1 isoforms were washed and boiled in 2 X SDS-PAGE sample buffers. PP1 inputs, Ddx21 eluates and negative control (-) eluates were separated and analysed by western blot for the presence of Ddx21 and PP1.</p

Topics: Biochemistry, Cell Biology, bacterially, pp1, isoforms, his-tagged, rna, helicase
Year: 2012
DOI identifier: 10.1371/journal.pone.0039510.g005
OAI identifier: oai:figshare.com:article/288144
Provided by: FigShare
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