Reprobing of western blot

Frederik Boernke boernke at ipk-gatersleben.de
Wed Jan 21 21:34:41 EST 1998


Hi all,

I tried to detect a GAL4 binding domain fusion protein in a western
blot using Clontech's monoclonal antibody. Protein extracts were
prepared
according to Printen and Spraque. Protein transfer was verified by 
staining with Ponceau S. Primary Ab was diluted 1:1000, secondary anti
mouse/HRP-conjugate was diluted 1:100000 (as recommended by the 
manufacturer). Signal was developed using Pierce's SuperSignal ULTRA.
But....I don't see any bands on the blot even no unspecific ones.
I thought I'd better repeat that with more "moderate" Ab dilutions.
Saving time, I'd like to reprobe the blot. Do you think I could use
it again without stripping (no signals anyway) or should I strip it?
Any stripping protocols available?

Thanks in advance
Ricky 

******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiologie
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
e-mail: boernke at ipk-gatersleben.de
http://www.ipk-gatersleben.de



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