ECL, ECF and Ab's

Bubba Zinetti zinetti at acpub.duke.edu
Sun Jun 15 14:44:31 EST 1997


I have been currently performing anti-phospho tyrosine western blots
from both cell lysate and from ip's.  I have been using HRP conjugated

4G10 (anti phospho-tyrosine) from upstate.  I now am interested in
actually quantifying some of the differences that I seen in my blots.
I have access to a Storm 680 phosphoimager, and I have done SOME
chemiflouresence with the machine.  My question is this, should I use
a FITC conjugated 4G10, or use the AP conjugated 4G10 for ECF
detection?  Obviously the FITC conjugated one would have the advantage
of less nonspecific binding and quicker results, BUT is it sensitive
enough?  Any input would be appreciated.  With the ECL detection I am
currently using some ip's need only 15-30 seconds exposure to give
great blots, while some others need 5-10 minutes.  I fear that I may
not see the results on some blots.
cya
BZ
Paul J Jansen                |Use Your Head
zinetti at acpub.duke.edu       |Sidestep the Traps 
http://www.duke.edu/~zinetti |Snake Through the Chaos
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