"Negative Staining" on Western Blots

HYDE at horta HYDE at horta
Tue Sep 28 08:26:31 EST 1993

In article <Berry.14.2CA7ECAC at biochem.unp.ac.za>, Berry at biochem.unp.ac.za (Ronald Berry) writes:
>Hello All
>Occasionally we have had a strange phenomenon with our Western Blots - 
>"Negative Staining". We get beautiful white bands on a pale (normal) 
>background, as well as the usual coloured bands. The chromogen is NBTS, and 
>the blots were blocked with milk powder.
>We haven't been able to pinpoint the cause of this, and wondered if anyone 
>else has had the same type of thing. Are these lipoproteins, perhaps? Why 
>else would a protein REPEL the chromogenic mix?
>If one can pinpopint the cause, it might be useful to do a kind of double 
>stain - some bands come out coloured, some white, on an intermediate 
>Ron Berry
>Biochemistry Dept
>University of Natal
>South Africa

I have seen this on some of my gels, too; the immediate cause I suspected was 
overloading the gel, and the large quantity of protein also transfers on the 
Westerns. How heavily are you loading your gels?
Fred Hyde				(Internet address pending)
Corporate Research Laboratories
3M Company
St. Paul, MN 

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