Detection of 3H-Leu by Autoradiography ?
zinc
zinc at zifi.genetics.utah.edu
Sat May 17 21:39:29 EST 1997
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scb1 at midway.uchicago.edu (Samuel C. Blackman) writes:
> 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.
There is a more simple method. Just soak the gel in 1M Na Salicylate
(Na salt of Salicylic acid) and then dry it down. If the gel is
higher than 9-10% acrylamide, soak it in some 5% glycerol to prevent
shattering. after drying it will be ready for autoradiography.
This technique is called fluorography, btw...
- -pjf
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