Smolke:Protocols/Western: Difference between revisions
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#Grow culture (between 5 and 25mL works well) in appropriate (generally dropout) media | #Grow culture (between 5 and 25mL works well) in appropriate (generally dropout) media | ||
#Pellet cells at 3000g, 4°C for 5 minutes | #Pellet cells at 3000g, 4°C for 5 minutes | ||
#*The idea is to lyse equal masses of cells in each tube. You need a large initial culture volume in order to measure the initial pellet mass. | |||
#Pour off supernatant, weigh, resuspend in water at 1 mg/uL | #Pour off supernatant, weigh, resuspend in water at 1 mg/uL | ||
#Transfer 100uL to a 1.5mL tube, repellet as before | #Transfer 100uL to a 1.5mL tube, repellet as before |
Revision as of 14:32, 21 July 2009
OverviewBlotting for large V5-tagged proteins in S. cerevisiae Materials
ProcedureLysis
SDS-PAGE
Semi-dry Transfer
Blotting
NotesReferencesContact |