[Histonet] RCC Antibody Help

[Sharon Chase]

 I am trying to establish a protocol for Vector's renal cell carcinoma
antibody. The data sheet states trypsin digestion for 30 minutes. I
obtain negative results with trypsin digestion using normal kidney and
clear cell carcinoma as controls. 
I then did HIER retrieval using Biocare's reveal and borg solutions in
their decloaker. The normal kidney's proximal tubules stained but the
clear cell was patchy. Repeating the run on the following day, using the
same reagents and working dilution, I obtained weak results with normal
kidney and negative tumor cells. 
A fresh daily working dilution(1/50), I can obtain constant normal
kidney staining but usually negative tumor cells.
I tried another tumor control block and leaving the antibody overnight.
 Normal kidney is fine.
Open to any suggestions.
 
Sharon Chase
Atlantic Health Science Center
Saint John NB
Canada