a high background of western - a high background of western (Apr/22/2011 )

I am doing the western blot.there is no problems when I transfer the proteins to PVDF menbrane.But I got a high background.

ken_zhang on Fri Apr 22 07:25:10 2011 said:

Hola, change the blocker agent or block for more time, because if you have background is because primary ab. bounds lightly but inespecifically to the membrane,I think. Buena suerte

ken_zhang on Fri Apr 22 07:25:10 2011 said:

Try blocking with 5% non-fat milk in PBS. Also dilute your antibodies in this. Also I've found that if you use your primary at too high a concentration you can get a lot of background. And wash LOADS! With plenty of wash buffer.