Hi CCPEM community,
I'm trying to solve the structure of a 140kDa complex (90+50) that has a relatively weak binding affinity (~2uM). I was going to start with negative stain, but I've found that at the low concentrations needed for negative stain, my complex falls apart. I've tried to crosslink it using GraFix and other methods, but have had very limited success crosslinking the complex without additional artifacts. This leaves me wanting to do negative stain at high protein concentrations (~2+ mg/ml). Does anyone know tricks for adsorbing FEWER particles to a grid so I can get away with using higher concentrations?
Thanks!
Erik
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