Guanidinium Chloride - (Jul/03/2009 )
do you use the same antibodies than your colleagues that succeed the stripping?
they must do something different. Do they use the same membranes (PVDF, nitrocellulose, from the same or from a different company)? I don't understand why you have black background, but the clue is in the little difference in the protocol of your colleagues.
-little mouse-
little mouse on Jul 21 2009, 12:31 AM said:
do you use the same antibodies than your colleagues that succeed the stripping?
they must do something different. Do they use the same membranes (PVDF, nitrocellulose, from the same or from a different company)? I don't understand why you have black background, but the clue is in the little difference in the protocol of your colleagues.
they must do something different. Do they use the same membranes (PVDF, nitrocellulose, from the same or from a different company)? I don't understand why you have black background, but the clue is in the little difference in the protocol of your colleagues.
We use the same PVDFs all of us. The only difference is that my Abs come in preserved in NaN3 whereas the others come in PBS. I really dont know if the NaN3 is making all the problem.
The day b4 yesterday i stripped and exposed to I Ab in 1:6000 concentration. All back. After that I stripped again, checked just with the secondary for signal (no signal so ok) and i exposed the blot to the same primary only the half concentration (1:12000).
Guess what... significant improvement...not perfect but conciderably better...
One other fellow did for me side by side the same with his membrane and my Abs...all black
So....what the h... is happening..is it possible that the NaN3 is causing te trouble? ANd if so, how can i avoid it? All my Abs are in NaN3
-Aris-