Running Retic lysate on SDS-PAGE

h ??? at vms2.macc.wisc.edu
Tue Mar 21 10:45:29 EST 1995



I have been doing translations in Rabbit retic lysate and am having a problem
with the translated material "sticking" at the interface of the stacking
and separating gels.  I'm using a standard SDS 1%, BME sample buffer
(laemmli).

I'm running with a 4% stacking gel and a 5-20% gradient gel with .1% SDS as a
final concentration..

I suspect the proteins being made are sticking to membranes left in the retic
lysate..  The RNA transalated is a full length picornaviral RNA producing
a `240 kDa polyprotein which is processed during and after the translation.
The sample buffer is added directly to the lysate and then boil for 5 min.

Thanx \
dave



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