Author: Blank, Maximilian F.; Chen, Sifan; Poetz, Fabian; Schnölzer, Martina; Voit, Renate; Grummt, Ingrid
Title: SIRT7-dependent deacetylation of CDK9 activates RNA polymerase II transcription Document date: 2017_3_17
ID: qm9urt2w_23
Snippet: The P-TEFb release assay was performed as previously described with some modifications (21, 22) . 5 g of anti-HEXIM1 antibody bound to Dynal magnetic beads (Life Technologies) were incubated with cell lysates (4 • C, 16 h). After washing, bead-bound P-TEFb/7SK snRNP complexes were incubated with purified Flag-SIRT7/WT or Flag-SIRT7/H187Y in the presence or absence of 2 mM NAD + for 2 h on ice. After sequestration of the beads with a magnetic se.....
Document: The P-TEFb release assay was performed as previously described with some modifications (21, 22) . 5 g of anti-HEXIM1 antibody bound to Dynal magnetic beads (Life Technologies) were incubated with cell lysates (4 • C, 16 h). After washing, bead-bound P-TEFb/7SK snRNP complexes were incubated with purified Flag-SIRT7/WT or Flag-SIRT7/H187Y in the presence or absence of 2 mM NAD + for 2 h on ice. After sequestration of the beads with a magnetic separator, the supernatants and bead-bound proteins were analyzed on western blots.
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