Personally, considering the low sensitivity of the direct method, I would just treat your antibody as if it where not conjugated, and do a species specific polymer or ABC/LSAB method. This way you can avoid Tyramide amplification unless really necessary....Like any new antibody, you will have to optimize your method to give you correct specificity and sensitivity...........
regards Michael Ho, MLT Resource Technologist Immunopathology laboratory Division of Pathology DPLM The Hospital for Sick Children Toronto, Ontario Canada Tel#:416-813-5950 Fax: 416-813-5974 Kim Merriam <kmerriam2...@yah oo.com> To Sent by: Histonet ih...@googlegroup <histonet@lists.utsouthwestern.edu> s.com , ih...@googlegroups.com cc 2009-05-21 08:41 Subject AM [IHCRG] HRP-labeled primary antibodies Please respond to kmerriam2...@yaho o.com Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet