In Dec. 1999, I posted a question concerning Ki67 staining in canine 
tissue. Thank you Jennifer MacDonald, Andi Kappeler, Mary Vaghan, and 
Gayle Callis for your responses. These responses, in addition to help 
from Maria in tech support at BioGenex, helped us to develope a 
reliable staining procedure that works well for our lab. We doubled 
the time in antigen retrieval (40 minutes instead of the usual 20 
minutes). We use a steamer and our usual citrate buffer. I feel 
certain that autoclaving or microwaving will work as well- we just 
happen to use a steamer. From what I understand increasing antigen 
retrieval time is the trick to getting this antibody to work. We use 
DAB as the chromagen and increased the time from 12 minutes to 15 
leaving the times throughout the rest of the protocal the same. If 
anyone needs further info I will be glad to share what I know. Thank 
you histonet for coming to the rescue once again!
Kathy
                Thank you 
 Kathy Oprea                     
 University of Georgia          
 College of Veterinary Medicine  
 Athens, GA 30602-7388           
 KOPREA@calc.vet.uga.edu         
The worker who's never said "Thank
God it's Friday," probably never
really worked.