"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 
>background.
>
>Regards,
>Ron Berry
>Biochemistry Dept
>University of Natal
>Pietermaritzburg
>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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