[Histonet] Pericyte, smooth muscle cell IHC problems

[Melissa Gonzalez]

Dear all, 
I have been trying unsuccessfully to stain for pericytes/ smooth muscle cells in sub cutaneous gliomas in a mouse model. (Tissues are either 4% PFA perfused or 10% FFPE)
I have tried several products, including:
* NG2 (Chemicon): works nicely in normal mouse organs, but binds to much of the tissue in the gliomas, making it impossible to interpret. 
*PDGFRB (Santa Cruz): resulted in a few crazy-appearing vascular areas, very weird
* ASMA (DAKO, Sigma, US Bio, Neomarkers, Abcam)
*Desmin (DAKO, Santa Cruz)
--in both monoclonals (using a mouse on mouse kit which did not work) and polyclonals, for which I only see staining in the attached skin and a very rare area in the tissue. I have tried both immunofluorescence on the perfused,  and chromogenic detection on paraffin sections, using Citrate, high pH, pepsin, and proteinase K retrievals. 
 
Is it possible that smooth muscle cells would not be well developed around blood vessels in a sub cutaneous glioma, and that is the reason I only see staining in the attached skin with a rare occurrence in the tumor? (Tumors are very vascular, blood vessels are definitely present)
 
Also, what is everyone using for blood vessel markers? We used to use BD Pharm. Pecam-1, which doesn't work anymore on anything except frozens, Santa Cruz goat Pecam-1 was working great but the most recent lots I ordered now appear completely negative using any detection. I have now switched to using CD105.
 
Any help would be greatly appreciated as I have tried all the tricks I know, and am left completely puzzled. 
 
Thanks so much, 
Melissa