Triton X-100

[Kyle Legate]

DK wrote:
> 
> 2. Even after filtering out the obvious insoluble light lipid-like 
> fraction, the resulting TX-100 lysates have almost magical 
> ability to clog columns. With large scale/concentrated
> lysate, it's very quickly getting to a point where the column 
> won't flow by gravity at all. None of that ever happens with 
> E.coli lysates. I always (perhaps naively) interpreted this
> as "cholesterol-rich lipid-protein complexes are not soluble 
> in TX-100 and get aggregated/concentrated instead, binding 
> to everything and cloging pores in agarose". 
> 
Insolubility in TX-100 is one of the hallmarks of so-called "lipid 
rafts", and very well might be the cause of clogged columns, but if the 
lipidic secretion the OP is working with is low in cholesterol, TX-100 
should definitely be considered in the screen. If the secretion is 
cholesterol rich, adding some digitonin might help to solubilize it.

> Octyl-glucoside would be my first choice to mix with 
> CHAPS if I must mix CHAPS with something. In some cases, 
> we also had pretty good results with LDAO (in terms of 
> rescuing difficult proteins, no 2DE). 
>