Re: [Histonet] LCA antibody for B5 tissue

From:"Bryan Hewlett"


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.


----- Original Message -----
From: "Moklebust, Amanda C" 
To: "'Patti Loykasek'" ; "histonet"

Sent: Wednesday, March 09, 2005 4:11 PM
Subject: RE: [Histonet] LCA antibody for B5 tissue


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.


-----Original Message-----
From: Patti Loykasek []
Sent: Wednesday, March 09, 2005 12:29 PM
To: histonet
Subject: Re: [Histonet] LCA antibody for B5 tissue

    Iım almost positive Iıve 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
Iıve 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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