I also recommend you to check Dan's link on the paper for stabilising
protein solubility.
I have similar experience. I got high yield, soluble protein but
aggregates once I have purified concentrated protein.
Before venturing to the detergent area, I would suggest using other
additives first (kosmotropes, chaotropes, amino acid, sugars and
alcohols). I managed to get it work in high NaCl, KCl and NaBr.
I managed to purify the protein, put it in Tris buffer with low NaCl.
I concentrated the protein and work fast enough before it crashes (my
protein usually crashes overnight). Then aliquot out the protein in
different eppendorf tubes containing different additives, leave it
overnight; the next day, I spun all the tubes and check for
aggregation (of course, make sure you leave a "control", which in my
case is Tris buffer, low NaCl; just for a point of comparison). I got
7 tubes then which do not have aggregated pellet; I left them for few
more days; 4 of them aggregated, and 3 of them looks okay.
Allan
Quoting Vitali Stanevich <stanev...@wisc.edu>:
Hi,
Sorry for off-topic question.
Does anyone have experience of the stabilisation of water-soluble proteins
by detergents? Protein I'm working with is definitely water-soluble and has
high yield, but, unfortunately, not very stable. Especially during
concentration. So, we thought that adding some detergents may one of the
ways to stabilise protein.
So, did anyone do it before or may be know published examples? Any
suggestions on the detergent type/concentration would be welcome.
Thanks,
Vitali
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