Dear cytometrists, I am trying to work up an immunohistochemical protocol trying to determine the presence of an enzyme with a double-labelled antibody system. The secondary Ab is labelled with Cy-3. My signals are very weak, even in the positive control. My basic question is this. Am I just dreaming that I will get a strong enough signal on individual cells without a signal amplification system? The protocol I am following suggests it can be done on cells adhered to slides, so I assume it is possible. Furthermore, I am kind of new to this and the protocol calls for rinsing after the secondary Ab incubation step. What should I be looking out for when it comes to avoiding washing out the antibodies. I have been consulting texts and they are some help but I thought discussing this with a live person would be more fruitful. I appreciate the expertise and hope I don't get chastised for asking a question that "could be" answered by a literature search. Thanks in advance. Steve Steven F. Mullen Graduate Fellow/Research Assistant Purdue University Department of Veterinary Clinical Sciences Cryobiology Laboratory Lynn Hall, G242 West Lafayette, Indiana 47907-1398 Phone: 765-494-0336 Fax: 765-496-1108 mullensf@vet.purdue.edu
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:30 EST