Help! Immunoscreen!!!

R. Sami Subbu sami at mail.wsu.edu
Tue Nov 25 17:48:06 EST 1997


Hi! 
I am screeing a expression library with an antibody. I have tried many
times but with no success! I have checked all the stages! Finally came to
a conclusion that there is no antigen on the filters. Because my positive
control where the purified protein is dot blotted lights up well! I am
trying to make it work for quite some time actually! I have done quite a
lot immunoscreen, but this time I dont know why I dont get any antigen
transferred to the filter! Every time I do the plaque lifting I get a
blank film! I have even probed with highly expressed antigen antibodies
but no signal! I have plated the phage in the presence of X-gal and IPTG 
I couldnt see any blue plaques! But the plaque morphology seem to be 
OK! Is there anyone who can help me in this case?  
Thank you!
-Sami Subbu





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