[Histonet] Vector Red and True Blue

[Ben]

Hello All,

 

I have been attempting some double labeling for fos and estrogen receptor
for about a month with little luck.  I am using Vector Red (fos) and True
Blue (ER) for my staining.  I am having no problems with the ER labeling,
but the fos labeling is not that clear.  I don't appear to be getting a
strong reaction product from the Vector Red.  Has anyone else had this
problem?  How strong should the staining be (light pink, dark pink, red)?  I
have been careful to only place in xylene for a few moments before
coverslipping and therefore don't think fading is my problem.

 

I have been thinking that my antigen retrieval may be the problem.  I am
using triton X to improve antigen retrieval, but was thinking about
microwaving.  These are free floating sections.  First is this possible to
do with free floating sections and second how long do I microwave?   Any
help for this lost soul is greatly appreciated.

 

Ben