Howdy folks!
I'm trying to crosslink a fairly large (approx 100kDa) protein to its
binding proteins on the cell surface (Bmax around 10e6). The problem
seems to be that the crosslinked products seem to be too large to enter
the stacker (4%) of the gel. I am using BS3 as the x-linker, (5mM,10m @25 C
in PBS).
I've tried shorter x-linking times (1min, awaiting results). I was
also thinking about subjecting x-linked products to some sort of enzymatic
or chemical degradation and then trying some sort of sequencing or something.
Anybody have any better ideas?
--
Jay Vasudevan
Dept. Pathology/Biochemistry
Box 214, UVa Health Sciences Center
Charlottesville VA 22903
