western blotting membranes - (Jun/27/2006 )
Hi, I have a stupid simple question about membranes in western blotting;
does the mambrane have a side when you transfer the proteins from the gel on it?
thanks a lot
I can tell you that PVDF and nitrocellulose don't
dnafactory is right:
it doesn't make any difference with nitrocellulose or PVDF which side you put onto the gel before transfer
but remember that after the transfer your proteins stick to one side of your membrane which means after transfer the side matters!
thank you very much, but when I have done some western without taking care of that even after the trasfer it still worked, how is it possible if they stick just to one side
thanks for the help
Id depends on the volume of Ab solution you use: if it is a lot and your membrane is somehow floating, the Ab will bind the proteins anyway. The major problem is when you develope the signal because you put the substrate only on one side. I always use the marker to know where my proteins are
and there are also pencils available to mark the membrane if you do not use prestained markers...
We don't spend too much to buy special things: we use normal pencils to mark the membrane. maybe in a place where we know we don't have proteins