Too clear bands in the centre of my WB
 

Hello everybody!

I’ve been having several issues with my westerns:

These days I’ve been detecting clearer bands in the centre of the membrane while the bands on the sides are ok... Even with the actins or GAPDH..!! Now I can't rely on my results... Does anybody know why is this happening to me?

http://img202.imageshack.us/img202/1052/wb1t.jpg

Re: Too clear bands in the centre of my WB
 

I had this problem for a while, and it was because my sandwiches were packed too tightly and were just "squeezed" better on the ends resulting in a more efficient transfer at the edges. Did you recently switch filter papers or sponges? That's when we caught it.

Are you using a stir bar in your transfer? That can be another source of what you're seeing - too much turbulence in the center. I no longer use the stir bar and have no problems with overheating.

Best of luck.