Detection of 3H-Leu by Autoradiography ?

Jonathan B. Marder marder at agri.huji.ac.il
Tue May 27 08:05:25 EST 1997


In article <EAAJA4.BHn at midway.uchicago.edu>,
   scb1 at midway.uchicago.edu (Samuel C. Blackman) wrote:

>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.)?
Fix the gel in  about 10% acetic acid (other fixers probably just as good),
wash the gel by shaking in water for 30 minutes (you can even just leave it 
under running tap water), shake in 20 gel volumes of 1 Molar Sodium Salicylate
for 1 hour, then dry directly. The reference for this is Chamberlain JP (1979) 
Analytical Biochemistry 98, 132-135.

Gels containing salicylate tend to take extra time to dry. You might want to 
have a layer of plastic wrap over the gel and leave it there during exposure 
to prevent sticking to the film.



Jonathan B. Marder             ,      Department of Agricultural Botany
E-mail: MARDER at agri.huji.ac.il |      The Hebrew University of Jerusalem
Phone: (08 or +9728) 9481918   | /\/  Faculty of Agriculture
Fax:   (08 or +9728) 9467763   |/  \  P.O.Box 12, Rehovot 76100, ISRAEL
     http://www.agri.huji.ac.il/~marder



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