Trichloroacetic acid

Protein isolation Bacteria - Bacillus subtilis

Experiment
Protein isolation Bacteria - Bacillus subtilis
Product
Trichloroacetic acid from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Protocol tips
- Protein extraction is performed following the publication protocol.

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‐PAGE

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Discussion

Discussion

5 years ago

Author: Hollie Fowler United Kingdom

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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Papers

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Trichloroacetic acid below.

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