Storing a protein membrane overnight? - (Mar/23/2005 )

Hi,

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?

Thanks

-cwong1215-

hi
you can incuber your membrane over a xeek end with your primary antibody that's no problem.

-fred_33-

Just in PBS @ 4C? With or without agitation?

-cwong1215-

QUOTE (cwong1215 @ Mar 23 2005, 01:40 PM)

Just in PBS @ 4C? With or without agitation?

I'd rather put it in TBS-T @ 4C, no agitation

Simon

-Simonsays-

hi,

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.

gud luck

-supriya-

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!

-bassamfahmawi-