Western help needed!

WZhao80850 wzhao80850 at aol.com
Fri Jun 30 23:56:56 EST 1995


Hello there,
I have big problem with my western blotting. I am using Amersham ECL
detection system. I use very high primary antibody dilution, i.e. 1:25
dilution, 1:1000 dilution with my sencond antibody. I always get very
faint bands, it seems like I miss the peak level of signal all time. Has
anybody dealt with low titer, high dilution primary antibody in western
blotting?
My target protein is about 27 kd protein.The bands I got on film are
always curved instead of straight bands. 
I am very frustrated here, any advice will be appreciated.
Thanx in advance!
Cheers
Wei
Wei Zhao
WZhao80850 at aol.com or weiz at protein.bchs.uh.edu
Tel: (713)669-1091(H), (713)790-4696
Baylor College of Medicine
Houston, TX 77030



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