[Histonet] EGFR

[Traphagan, Patricia]

We have been trying to achieve good staining using this antibody. We use
the Dako EGFR kit. Dako's control that comes with the kit, turns out
beautifully, which tells us that the stain procedure is correct. Our
problem lies with our internal control. The pathologist are just not
happy with it. According to them we have: Over staining of the smooth
muscle, normal epithelial tissue does not stain correctly, and have
un-uniform staining of the membranes. Can anyone tell me what this
means? We use 10% NBF, fixation is probably no more than 8 hours. Our
processing run is about 12 hours for large tissue specimens, and we use
an alcoholic formalin in the second station, which then goes to 70% ETOH
and the graded series of alcohols, xylenes, paraffin. We do not put the
cut tissue in the oven, we let it air-dry overnight, possibly two
nights. We are at a total loss. Any suggestions. We really need
HELP!!!!!