crosslinking in organic buffers

Thierry Baussant tbaussan at club-internet.fr
Wed Jun 11 14:48:47 EST 2003

Try NHSbromoacetate (see Pierce)

Thierry Baussant PhD

"Scott Craig" <s.craig at student.unsw.edu.au> a écrit dans le message de news:
3EC98196.30705 at student.unsw.edu.au...
> Hi All,
> We are trying to couple a sulfhydryl labelled peptide to a lipid
> containing free amine groups. So far we have attempted to do this
> reaction in a two step process by reacting the NHS group of the LC-SMCC
> (a NHS and maleimide containing cross-linker) with the amines on the
> lipid in a buffer of DMSO+50uM triethylamine followed by reverse phase
> HPLC seperation and confirmation of the size using MS. This reaction
> appears to work from the MS results, however when we have subsequently
> tried to couple the maleimide group with the sulfhydryl group we appear
> to not get any coupling as analysed by HPLC. The maleimide/sulfhydryl
> labelling was done in 50% ethanol+50% HEPES+1mM EDTA. We have  used the
> LC-SMCC to fluorescently label a protein in aqueous buffers to confirm
> the reagent is working. Does anyone have any suggestions on how we could
> go about doing this crosslinking reaction in organic buffers or any
> other suggestions.
> Thanks for any help.
> --
> Scott Craig
> Biotechnology & Biomolecular Sciences
> University of New South Wales
> Sydney 2052, Australia

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