Storing a protein membrane overnight? - (Mar/23/2005 )
Quick question. I ran a western blot earlier today and realized that we had a secondary antibody that was isotope specific (non-compatable with what i was doing). So I put in an order for a new secondary which I hope will be in tomorrow. My question is which route should I take with my membrane that has already been incubated with primary antibody (anti-phospho p27)
Should I A) strip and reprobe and hope that the antibody comes in tomorrow or store the membrane overnight in PBS @ 4C.
My main concern about is whether I will have a huge amount of signal degradation. Has anyone had positive experiences with storing their membrane overnight? If so, what method did you use?
you can incuber your membrane over a xeek end with your primary antibody that's no problem.
Just in PBS @ 4C? With or without agitation?
I'd rather put it in TBS-T @ 4C, no agitation
was just browsing through the forum topics and saw ur question. though u have put up this question quite a long time ago but still i would strongly suggest that u should never ever use PBS when ur detecting phosphorylation.
I have always kept mine over the weekend submerged in MiliQ water with 0.5% sodium azide without agitation at 4C over the weekend and had no problems. Good Luck!