IndraLab

Statements

USP29 binds GST. 8 / 8
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"In addition, purified GST‐USP29 but not GST could interact with His‐CDK1 in vitro, indicating the direct interaction between USP29 and CDK1 (Figure  xref )."
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"Bacterial‐expressed GST and GST‐USP29 protein bound to Pierce Glutathione Agarose was incubated with PET28A‐TWIST1, purified from the Escherichia coli strain BL21 overnight at 4 °C. Then the beads were washed with PBS four times, followed by western blotting."
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"Figure  xref illustrates that Flag‐tagged TAK1 in cell lysate could be specifically bound and eluted by GST‐USP29 fusion protein but not GST alone."
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"In addition, purified GST‐USP29 but not GST pulled down His‐TWIST1 in vitro, indicating the direct interaction between USP29 and TWIST1 (Figure  xref )."
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"The ubiquitinated TWIST1 protein was then incubated with purified GST‐USP29 WT and C294S protein separately for 4 h at 4 °C, followed by western blotting analysis."
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"The recombinant GST‐USP29 WT and 3A mutant protein were expressed in Escherichia coli strain BL21 and purified using Pierce Glutathione Agarose."
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"Next, Co-IP experiments revealed a direct interaction between USP29 and TAK1 (Fig. xref ), which was further confirmed by GST pull-down assays (Fig. xref )."
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"Purified CDK1 beads were finally incubated with purified GST‐USP29 WT and 3A in kinase buffer (50 m m Tris‐HCl, pH 7.4, 50 m m NaCl, 10 m m MgCl 2 , 10 mm β ‐glycerophosphate, 1 mm dithiothreitol (DTT), and 100 µ m ATP)."
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