Biotin/streptavidin/blocking

Gary Krause gkrause at cms.cc.wayne.edu
Tue Dec 10 13:49:09 EST 1996


Dr. David Martin wrote:
> 
> In article <199612051520.JAA05199 at borcim.wustl.edu>,
>    brett at BORCIM.WUSTL.EDU (brett) wrote:
> +>>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?

> 
> 0.25% gelatin in 50mM tris pH 7.4 150mM NaCl 5mM EDTA .1% Triton X-100
> also works fine. Another thing I find helps is to wash twice without
> detergent/block just prior to adding the ECL reagent.
> 
> 
> * David Martin - Atherosclerosis and Thrombosis research group  *
 


Does gelatin contain phosphorylated proteins?

-- 

Gary Krause (gkrause at cms.cc.wayne.edu)
Department of Emergency Medicine
Wayne State University, Detroit, MI USA



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