32P gel developping

Rogier rogier666NOroSPAM at hotmail.com.invalid
Fri Feb 18 13:10:02 EST 2000


Hi Paola,

>develop gel containing proteins labeled with 32P
>X-Ray films and intensifying screens

any type of X-ray film (try kodak X-Omat or Fuli X-ray film) and
an intensifying screen will do.

>thought to dry the gel before develop it

I would only do this if your signal is weak and you need long
exposure times. drying gels is always a gamble. sometimes they
crack during drying. If you really need to dry them (low signal,
many bands close together, ...) use a vacuum dryer. or simply
blot your gel onto nitrocellulose, then use the blot for
autoradiography/phosphorimaging.

>freeze the gel(-20 C or -70 C ?)

if you're gona use an intensifying screen, you'll need to freeze
the gel, 'cos these screens only work at -70C or lower. You'll
need to dry or blot the gel then, because freezing will kill
your gel.

>If I freeze the gel, could I also use it for immoblot?

you'll need to dry the gel before freezing. once dried, you can't
blot it anymore. I'd suggest blotting it and do autoradiography
and immunostaining on the blot:
- blot your gel, then block it with BSA, milk, boehringer
blocking reagent, or whatever blocking agent available.
- expose a few films with your blot, or use a phosphorimager
- then add antibodies, just like on a normal western blot. you
can do ECL. the exposure times normally used for ECL or much too
short to pick up the radioactive signal.

Good luck,
Rogier Stuger
E rogier at biogate.com


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