I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer.
Please assist me to select most suitable buffer and the concentration
TCA is a strong acid, so the best resuspension buffer is Tris-pH 10, with 2% SDS.
Dear Mr. Oscar,
Thank you for the knowledge sharing. It works well.
If you could please provide references.
No need for references. Follow the logic. Precipitation carried out with a strong acid like TCA, therefore precipitate is acid. You need to bring it into solution. There's nothing better than some electrical equilibrium by neutralizing charges = use opposite pH (alkaline). But pH has to be maintained, therefore you need a buffer. Which buffer? A good organic, popular, cheap and easily available buffer like Tris, HEPES or MOPS. What if the protein doesn't solubilize? Use a detergent. 2% SDS is a fairly good detergent. That's it I think.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts