[Histonet] LCA antibody for B5 tissue

[Bryan Hewlett]

Amanda,

We have used Dako's  M0701, clones (2B11 + PD7/26) on B5 fixed material for
lo' these many years with NO trouble.
It's even mentioned on the product data sheet.
HIER is not required! However, you should treat B5-fixed material with the
iodine/hypo sequence to remove mercury fixation pigments(even if you don't
see them).
Dilution=1:100 incubate 60 min @ RT.

Bryan

----- Original Message -----
From: "Moklebust, Amanda C" <amoklebu <@t> seattlecca.org>
To: "'Patti Loykasek'" <ploykasek <@t> phenopath.com>; "histonet"
<histonet <@t> pathology.swmed.edu>
Sent: Wednesday, March 09, 2005 4:11 PM
Subject: RE: [Histonet] LCA antibody for B5 tissue


Patti,

I have been pretreating the slides with Dako's Target Retrival
Solution pH 6.1 in a steamer for 20 minutes.  The slides do not get treated
to remove mercury pigment.  I've gotten two replies recommending a different
clone (2B11/PD7/26) from Dako.  I don't think they were posted to everyone.
Thanks for the help.

Amanda


-----Original Message-----
From: Patti Loykasek [mailto:ploykasek <@t> phenopath.com]
Sent: Wednesday, March 09, 2005 12:29 PM
To: histonet
Subject: Re: [Histonet] LCA antibody for B5 tissue


Amanda,
    I¹m almost positive I¹ve had this antibody work on B5 fixed tissues,
mainly bone marrows. What type of antigen retrieval are you using? This
could make a difference. Are you doing any type of treatment to remove the
precipitated mercuric chloride? This can lead to false negatives with some
antibodies.
I¹ve routinely used a citrate based heat pretreatment. This recovers the
antigen nicely and removes the mercuric chloride(in most cases).
Hope this helps.

Patti Loykasek
PhenoPath Laboratories
Seattle, WA>


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