[Histonet] Temperatures for IHC

[Ronald P. Wilson]

We are in the process of optimizing our IHC procedures. We have switched to
using some of the reagents from Phoenix Biotechnologies. Their recommended
protocols suggest primary and secondary antibody as well as strepavidin and
chromagen incubations be done at 55ºC for much shorter time periods (4-10
minutes). I have also heard that some labs do their incubations at 37ºC. Is
it common practice to use temperatures higher than room temperature? Are
there advantages other than just shortening the incubation time? If anyone
is using the higher temperature, how do you do this for bench-top
procedures? I know that some of the staining stations such as Microprobe
have built-in incubation chambers to do this.

Ronald P. Wilson, V.M.D., M.S.
Associate Professor
Department of Comparative Medicine, HO54
Penn State University College of Medicine
M. S. Hershey Medical Center
500 University Drive
Hershey, PA 17033
phone:  717-531-8460
fax:       717-531-5001
e-mail:  rpw4 <@t> psu.edu