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
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