IHC Related Problem

Nick Theodorakis via methods%40net.bio.net (by nick.theodorakis from gmail.com)
Thu Dec 20 10:36:30 EST 2007


On Dec 17, 10:23 am, "Sudheendra Rao N R" <sudhe... from gmail.com> wrote:
> Hello all,
> We are trying to do IHC on free floating 30 microns rat brain sections.
> Primary is antiGFAP (Mouse Monoclonal-1:1000) and secondary is antiMouse
> (Horse: 1:250), by ABC-DAB method.
> But we are getting reproducible nonspecific staining in SPECIFIC regions of
> the rat brain

[...]

> We did a negative control by skipping the primary..but same results..the
> cells seem to be of pyramidal / immature neuron morphology.
> We also skipped both primary and secondary, going ahead with ABC-DAB but no
> staining (rules out endogenous biotin).

Is your secondary (anti-mouse) antiserum guaranteed to be free of
cross-reactivity to rat antibodies?

Nick

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Nick Theodorakis
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