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文献和实验7 or SP6 Kit from PROMEGA Glutathion-Agarose beads from Pharmacia Protocol: a) Resin preparation (50% slurry): 1- Transfer a 1.33 mL aliquot of the resin (mix well before use) into a 15 mL Falcon tube 2- Centrifuge (1pulse at 500g) 3- Resuspend in 10
GST Fusion Protein Preparation
debris at 10K 10' 4℃.Aliquot supernatant and store at -70℃.BINDING TO BEADS AND THROMBIN CLEAVAGEIncubate soluble lysate with 1/2 volume of 50% GST beads at 4℃ rocking on the Nutator for 30-60 minutes.50 ml conical Falcon tubes work well.Pour
with 1/2 volume of 50% GST beads at 4C rocking on the Nutator for 30-60 minutes. 50 ml conical Falcon tubes work well. Pour into a BioRad Column and drain lysate. Save 100 ul of depleted lysate. Load 2.5 ul for SDS-PAGE. Wash beads with 3 column volumes
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