Reducing non-specific binding in a radioligand binding assay - (May/26/2008 )

Howdy.

I was wondering if there was a specific method/procedure one can use to reduce non-specific binding in a radioligand binding assay. I'm looking at the binding of [3H]-methyllycaconitine to nicotinic ACh receptors embedded in the membrane of T-cells.

I have tried many different buffers. The only common aspect between them seems to be Tris-HCl and a certain w/v of BSA. My best results seem to be with my most simple buffer - PBS + 0.1% BSA. But the non-specific binding is still very prominent. Any suggestions? Or should just continue to plough through the literature and try random buffers suited to [3H]-MLA binding?

Any feedback would be greatly appreciated !