Biotin/streptavidin/blocking

brett brett at BORCIM.WUSTL.EDU
Thu Dec 5 10:21:54 EST 1996


>Fellow researchers:
>I've boitinylated cultured cells, lysed with 2X SDS gel loading buffer with 
>BME, sheared DNA, ran on SDS PAGE, transfered to PVDF, blocked with 5% 
>milk/TBS+0.05% tween 20, probed with streptavidin-HRP conjugate and 
>detected with ECL detection solutions 1+2, home made but good:-)
>I would like to know what is the best blocking solution for this procedure?
>Thanks, Mike
>-- 
>Mike Garbutt,
>Dept. of Anatomy and Cell Biology,
>6-32B Medical Science Building,
>University of Alberta,
>Edmonton, AB.
>Canada.T6G 2M7
>Tel. (403) 492 6485 (Work)
>Tel. (403) 465 9285 (Residence)
>e-mail:   mgarbutt at gpu.srv.ualberta.ca
>homepage: http://www.ualberta.ca/~mgarbutt/

I've used 3% BSA *or* 5% nonfat dry milk in TBS-tween, with equivalent results.
The BSA costs more, though.




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