Actin + another protein - (Jul/17/2007 )

How can I do to have in the same blot and sample the Actin and another protein?
I incubate the both Ab in the same time? The MW of the Actin is?

Usually beta-actin is so strong that I strip the membrane and re-incubate

I would not recommend incubate both 1st Abs together in any case, especially when you are using either Ab the first time (see http://www.protocol-online.org/forums/inde...showtopic=28791 for the reasons). If you know their sizes and they are reasonably apart, you can cut the membrane into two and probe each with relevant 1st and 2nd Abs. If that is not possible and you have enough samples, you can run two sets (on the same gel if possible) and probe each set with one 1st Ab. Else you can strip the membrane (if the sizes are too close or the 1st Abs are from the same species) or using azide to quench the 2nd Ab HRP (the 1st Abs have to be of different species) and then reprobe (if you use azide you have to wash carefully before reprobing)