Stripping membrane - (Sep/26/2006 )

Hi guys,

I have a question for you. For the moment I run a lot's of western blot and I need to blot the same membrane with different antibody. The different antibody have almost the same size. So I strip my membrane after use. Do you know how many time can I strip a membrane????

Thanks a lot

Biofred

You mean how many times you can reprobe your membrane, isn´t it?
I think that between 3-4 times you can get good results, although you should try yourself.

it depends, among others, on the kind of antibodies_; if you use first , let´s say, an antirabbit IgG, then an anti-mouse, and then an anti-goat, you even may pass on stripping (but may do this), and simply inactivates HRP secondary antibody by NaN3; if primary antibody is frome same species, problems may increase with multiple stripping step; start with antibody of anticipated lowest signal; I think there is no general rule, as it depends on stripping protocol and strongness of signal; suggested 3 to 4 times stripping may succeed;

follow also the discussions upon stripping protocols

Thanks a lot guys for your answers

Have a nice day

Biofred