western blotting need help can i not strip if using same ab - (Jul/06/2006 )

HI, i got no signal on a western blot I currently did because i used the wrong secondary antibody, do i have to strip the membrane or can i just block as regular and do it again. i am looking at proteins such as s6, akt, thanks

-titans-

QUOTE (titans @ Jul 6 2006, 06:06 PM)

what is wrong in your secondary antibody?
Does it bind on the same epitope than the right antibody? if yes ; strip, if not, no need to strip.
please give more information, we cannot help.

-Missele-

thanks for the help

i used goat anti-rabbit and i needed goat anti rat, would that bind and cause a need for stripping?

-titans-

If u used anti-rabbit instead of anti-mouse, just incubate in the right antibody again and it should work without stripping.

As Missele suggested, more info would b better.

-scolix-

thanks for the help guys,
i think i got it, im just applying the right antibody on it now,

-titans-