Backgound problems in Western blotting - HELP!

[ijiwaru at wheel.dcn.davis.ca.us]

In article <4of00m$uh at underworld.centrum.dk>, mgl5403 at centrum.dk (Med.
Gas. Lab.) wrote:

> Hi netters
> 
>    I am doing Western blotting and using the ECL-detection system. My
> problem is that I gt an irregular, uneven background. I have tried
> modifying washing procedures and blocking buffers.
>    I am using the PHAST-system from Pharmacia. Does anyone out there
> have an absolutely super protocol, that I can try out??
> 
> Greetings
> 
> Jakob Hendel
Jakob,

I've being doing westerns using NEN's Renaissance reagent, very similar to
the Amersham ECL.  I've been rinsing the blot in tris-buffered saline
after incubation with the chemiluminescence reagent before exposure of the
film.  It seems to help with the background, IMHO.  I don't know how well
it will or will not work with PHAST gels, YMMV.

Lyle Najita
Plant Pathology
University of California - Davis