Protein immobilization on nitrocellulose membrane - (Sep/07/2006 )

Hello!

I have to immobilize cell lysate on a nitrocellulose membrane for a dot blot, but I don't know how to do it. In the reference's papers I read they just say "lysate was immobilized onto nitrocellulose", but never explain the procedure.

Could somebody help me?

Thanks in advance!

you pipet a small drop of sample onto the nitrocellulose membrane (which is either supported by absorbant paper or on a vacuum blotting device). the liquid passes through the pores and the protein binds to the membrane.

don't forget to wet the membrane to prefill the pores, just float the membrane in a dish or tray of water (getting rid of bubbles underneath) until the water comes through the pores.