I have been using 2% Triton X-100 followed by 20% sucrose in my cell wash buffer post cell lysis. Triton being a mild detergent, I would like to know the use of sucrose apart from its role in acting as a denser medium in density-gradient centrifugation during cell fractionation.
Compounds like sucrose will increase the stability of proteins in the lysate, by lowering the water activity. I have also seen claims that sugars may stabilize lysosomal membranes, thus preventing protease release https://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/lysis_buffer_additives/
BR Birger
Thanx a lot.... I would also like to know if the use of sucrose in such a buffer is concomitantly significant for IB proteins only. If my protein is expressed in the soluble fraction, does it work similarly?
The stabilization should certainly apply to the soluble fraction..I suppose (but this is just guessing) that it the lowering of the water activity will also inhibit protein aggregation and adhesion to surfaces..
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