[Histonet] re: phosphate buffered saline or Tris PBS ?
|From:||"PW Howorth, Physiology" |
I'm currently using immunohistochemistry (free floating sections or
serially mounted sections) to map the rat brain. We use viral injections
to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3
or AMCA) to colocalise receptors with GFP expression.
I have been trying to improve our methods that we currently use.
Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups use
a variety of differences in these recipes, what advantages does Tris PBS
offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ?
Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction
with GFP signal compared to just using 50% ethanol ?
And finally, we use cardiac perfusion to initially fix tissue followed by 2
hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin.
The antibody we use for mu opioid receptors (diasorin) - does anyone have
any experience with this antibody and this method have you done antigen
retrieval to improve the signal ?
Dept of Physiology,
University of Bristol,
+44 (0)117 33 17112
Histonet mailing list
<< Previous Message | Next Message >>