boiling membranes - (Apr/28/2008 )

Hi everyone,
I am having trouble expressing a specific signal with my antibody (a small cytokine) and I heard that it might be good to boil the membrane in PBS before the blocking step. Does anyone know the theory behind boiling the membrane?
Thanks in advance

Hi, i never hear about boiling membrane,
anyway, i think that boiling could expose better (or in different way) your protein epitope, so that could be seen better by your Ab..

hey...
I have never done that to my menbranes; but, when people want to mark ubiquitin or ub-proteins it is a usual step to autoclavate the membrane, to ennhance the specificyti/signal; Try to search the web with "membrane autoclavate" i am sure you'll find soem information.

thanks, i'll take a look!

Yes, autoclave is necessary for ubiquitin WB and it works very good in our lab.