sulphoEGS protein cross-linker

EK nobody at elnino.com
Fri Jul 11 13:37:43 EST 2003


"Szymon Z." <szymon at nospam.please> wrote in message
news:bemjkj$eoj$1 at atlantis.news.tpi.pl...
> I am attempting to do a series of protein - protein cross-linking
> experiments using Pierce Sulpho-EGS reagent.
> The booklet says, that EGS solution should be prepared just before use,
and
> that EGS (in substance) vial should be dessicated for it is hydrolysed
> easily.
>
> My question is, how fresh the solution shall be - will it withstand a few
> hours, or a few minutes before loosing efficiency or reproducibility?
> Or maybe I may freeze it in nitrogen in aliquots and use them for a few
> days?
> Will frequent opening of EGS vial will deteriorate its quality very
rapidly?
>
> Any practical advice will help me very much in planning and conducting
> experiments, as in my lab we have never used that EGS before.
>
> Thanks a lot
>
>   - Szymon Zietkiewicz
> szymon @ biotech univ gda pl
>
>

I am almost sure that you should work very fast after reconstituting the
reagent in the reaction buffer. 10 min will be too long. I would even say 5
min. You still might need to run the rection to completeness, and that may
require 30-60 min incubations, sometimes even several hours, but first 5
minutes are most important. You certainly should contact Pierce for details,
but my experience with similar applications involving succinimide esters
sais that the key is in doing everything very quickly. Also, prepare only
enough reagents for one reaction at a time and discard leftovers.
-Emir





More information about the Methods mailing list