3 replies to this topic

[cwong1215]

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

[fred_33]

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

[cwong1215]

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

[Simonsays]

cwong1215, on Mar 23 2005, 01:40 PM, said:

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

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

Simon