I have seen this done many ways, some people have h202 at th= e end of their depara setup if doing that off line. For the Leica Bon= d instrument we were advised to do h202 block just before the DAB and i hav= e noticed that it works best that way for their system. I for years h= ave always used h202 after the primary ab as some abs are affected by it so= I just don't take the chance of not knowing which may be. There was = one ab, cd4 I believe that for whatever reason needed to be h202 blocked be= fore AR and the AB, and it really didn't work if you did not do the block b= efore everything. Just goes to show that each ab has to be handled sp= ecifically.
A little aside here and techni= cally it makes no sense but we were having trouble with the Leica ap/red ki= t preciptating and was advised by Jim Burchette not Leica to use H202 = just before the red substrate chromogen and that has really improved the ap= /red staining for us which we use a lot on derm samples. I know = endogenous alk phos uses levamisol as a block and not h202 but for some rea= son this works so I don't argue with it. R= egards, Patsy -------- Original Message --------Subject: [Histonet] Immuno. confusion From: "Hannen, Valerie" <valerie.han...@parrishmed.com<= /a>> Date: Wed, August 03, 2011 8:57 am To: "[1]histonet@lists.utsouthwestern.edu" &= lt;[2]histo...@lists.utso= uthwestern.edu> Hi folks... I am going to try to revam= p out Immuno. procedure and am looking at a couple of spec. sheets from our= primary vendor. I am gettin conflicting information...so I am turning to y= ou all for help. When doing immuno's that require either HIER or enz= yme digestion, do you perform the retrieval first or do you do the "peroxid= e" step first?? One spec. sheets says retrieve then block endogenous per oxidase...the other says block endogenous peroxidase then do the HIER or en= zyme digestion..which is correct?? One other question that I have is= ...what do you use to enhance your stain?? The enhancing solution that we w= ere using has been discontinued. Thanks so much!! Valerie Han= nen,MLT(ASCP), HTL,SU(FL) Histotechnologist Parrish Medical Center 951 N. Washington Avenue Titusville, Florida 32796 (321) 268-6111 ex= t. 7506 ****************************************************= ********** "This email is intended solely for the use of the individual = to whom it is addressed and may contain information that is privilege= d, confidential or otherwise exempt from disclosure under applicable law= . If the reader of this email is not the intended recipient or the emplo= yee or agent responsible for delivering the message to the intended reci= pient, you are hereby notified that any dissemination, distribution, or<= BR>copying of this communication is strictly prohibited. If you have rec= eived this communication in error, please immediately delete this messag= e. Thank you" **********************************************************= **** _______________________________________________ Histonet mailing= list [3]Histonet@list= s.utsouthwestern.edu [4]http://lists.utsouthwestern.edu/mailman/listinfo/his= tonet References 1. 3D"mailto:his 2. 3D"mailto:histonet@lists.utsouthwestern.edu" 3. 3D"mailto:Histonet@lists.utsouthwestern.edu" 4. 3D"http://lists.utsouthwestern.edu/mail_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet