Trichloroacetic acid
Protein isolation Bacteria - Bacillus subtilis
Publication protocol
The culture supernatant was collected and separated from cell fraction by centrifugation at 5000 g for 30 min at 4°C with iterative removal of the supernatant into a fresh tube for 4 rounds. 40ml of the clarified supernatant was precipitated overnight with 6.25% (w/v) trichloroacetic acid (SigmaTM) at 4°C and the precipitated proteins were recovered by centrifugation as before. The protein pellet was washed 5 times with 1ml ice‐cold dH2O and air dried for 1h. The protein pellet was suspended in 50 µl 2x Laemmli buffer and separated by SDS‐PAGEFull paper Login or join for free to view the full paper.
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Discussion
Discussion
5 years ago
Author:
Hollie Fowler
How can I deal with my pellet being too viscous?
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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Manufacturer protocol
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