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2 years ago
2 years ago by 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?
Found 2 matching solutions for this experiment
|- Cells are removed by centrifugation at 8,000 × g for 10 min. The supernatants are retained and sterilized by filtration through a 0.22-μm-pore-size Millex GP filter (Millipore, Billerica, MA).|
|- 20 ml of culture supernatants are precipitated by the addition of 100% (wt/vol) trichloroacetic acid (TCA) at −20°C to a final TCA concentration of 10% (incubated on ice for 45 min before centrifugation at 4,500 × g for 45 min at 4°C). 3 ml of ice-cold acetone is added to the pellet (15min before centrifugation at 4,500 × g for 45 min). The pellet is air dried before resuspension in 130 μl of rehydration buffer.|