Detection of 3H-Leu by Autoradiography ?

Samuel C. Blackman scb1 at midway.uchicago.edu
Fri May 16 15:22:51 EST 1997


I've been labelling proteins with 3H-Leu, and have been quantitating
changes in protein levels by liquid scintillation counting.  I am curious,
though, whether or not I can resolve these proteins by SDS/PAGE, transfer
to nitrocellulose or PVDF and then detect the labelled proteins by
autoradiography.

I've heard that it is possible if you spray the membrane with a liquid
scintillant spray.

My questions are:

 a.) Does any one do this and have protocol for it?
 b.) Does anyone have any idea about what brand/type of spray to use?
 c.) Can anyone provide any technical information (such as the minimum
     specific activity of my 3H-Leu needed, exposure time, conditions,
     etc.)?

Any and all help would be, of course, greatly appreciated!

Thanks!

- Sam


-- 
Samuel C. Blackman        ! InterNet : blackman at tigger.uic.edu
MD/PhD Student (4/8)      ! Disclaimer: I speak for me, not UIC!
Univ. of Ill. at Chicago  ! Quote : "Quandro potro io finir di stupire?"
Dept. of Pharmacology     ! Phone : 312/996-4983 (lab)  Fax: 312/996-1225



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