Hypoxyprobe - Anyone used is Successfully over long term? (Jan/24/2008 )
Need some help with hypoxyprobe treated tissue. It turned out great after a week once it was embedded. But trying it again after 5 months, and signal is very very low.
Anyone has any experience with the probe? Please help.
It is used to detect hypoxia in tissue/tumors. Apparently the company got bought out and doesnt sell it anymore, and obviously has miminal tech support for it.
What is your embedding medium ? Is your probe fluorescent ? How were conserved your slides ?
The probe itself is not fl. It is injected into the animal with a half life of 4 hours. I sacrificed the animal after 2, and collected the tissue, and fixed it in formalin. It was then embedded in paraffin within 2 days, and the first set of staining done in a week. It has a primany antibody which comes with the kit and a biotinylated secondary, and then a DAB detection.
Hope this helps. At this point any input is greatly appreciated
After DAB staining, how did you mount your samples ? Permanent or aqueous medium ? What do you use for counter-staining ? Is your DAB stable in alcohol ?
if yes, you'd rather counterstain with alocholic Hematoxylin, then seal your slides with permanent mounting medium such as entellan, and keep your slides in the dark.