cross-linking

LOWDEN,Darren BOTDJL at LURE.LATROBE.EDU.AU
Sun Oct 11 19:27:43 EST 1992


Dear fellow netters,

I am interested in trying to identify the cell surface receptor to which a
protein of interest binds to. I am using a chimaeric protein (fused with the
maltose binding protein) so I thought that one means of accomplishing this task
would be to cross-link the chimaeric protein in the presence of cells,
solubilise the cell membranes and pass the entire extract over an amylose
column in the hope that the maltose binding protein tag will bind. By washing
the column and then eluting bound material and reversing the cross-linking
process I hope to be able to resolve two bands on a SDS-PAGE gel, one band 
being the chimaeric protein and the other hopefully the receptor of interest.

Unfortunately I have no experience whatsoever with cross-linking reagents and
would welcome any advice with regards to reviews, references, protocols,
companies that supply reversible cross-linkers etc.

Thanks in advance for any help that you may be able to offer.

Yours sincerely,
Darren Lowen
botdjl at lure.latrobe.edu.au



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