|- Bacterial pellets are recovered after centrifugation (10,000rpm, 10 min), washed twice in PBS, and resuspended in 100 µl of B-PER II bacterial protein extraction reagent following the manufacturer's protocol.
|- Culture supernatants are precipitated in 60% ammonium sulfate or trichloroacetic acid at 4°C overnight. After centrifugation, pellets are washed twice with 5ml cold acetone, dried, and resuspended in 200 µl of 2× Laemmli buffer.
B-PER™ Bacterial Protein Extraction Reagent from Thermo Fisher Scientific has not yet been reviewed for this experiment
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4 years ago
Author: Hollie Fowler
I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?
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