|- Cell cultures centrifuged at 14,000 rpm for 5 min.
- Supernatants were passed through an 0.2-μm Acrodisc syringe filter.
|- Cell pellets are resuspended in 100 μl of B-PER protein extraction reagent and incubated for 10 min at room temperature. After, they are centrifuged at 14,000 rpm for 10 min, and supernatants are recovered.
- If there are insoluble precipitates (from membrane fractions) they can be resuspended with 50 μl of B-PER protein extraction reagent.
B-PER™ II Bacterial Protein Extraction Reagent (2X) 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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