ttha at uhura.cc.rochester.edu (Tom Thatcher) wrote:
>I want to run an SDS gel and Western blot on mouse liver. I have
>2 strains of transgenic mouse (made by others) and I would like
>to verify that they are expressing the transgene. One is a
>metallothienin promoter, the other is MHC class-I, whihc should
>be nearly ubiquitious. Is there any reason I can't just drop a
>piece of liver into SDS-PAGE loading buffer and boil it?
>If so, what should I do instead?
At least I would homogenise the tissue first. It may also pay to do a
crude separation into different fractions by centrifugation (depending on
were your protein is). That way you reduce the amount of irrelevant
proteins and therefore detection sensitivity.