reprobing old membrane without stripping - (Jul/25/2006 )
i tried to reprobe old membrane kept at 4C for months in saran wrap without stripping. i can't see the band and the background is high (some spots). I am wondering could it be because i didnt strip it? thanx
sorry the membrane was kept at 4C in PBS-T for around one month.
well there is a possibility that not stripping is the cause of your problem. I store membranes without stripping but strip them before reprobing (doesn't stink your membrane after 1 month?...)
thank you Fred, it is really tricky cause stripping might cause damage to the protein right? and no stripping doesnt make my membrane clear... ...but i guess since it's first time we are trying this antibody and optimizing the conditions it is better to strip ...maybe
for optimising a new Ab, i would use a fresh blot.....just to remove any possible probs with the strip...#
as for damage to thr protein from stripping - a possibility, but has never been a problem for me.....my labs srtips routinely up to 4 times....
stripping can reduce the signal, that's why you should always label first with the lowest affinity antibody.
aussieuk is right, do your optimization on a fresh blot.