Re: [Histonet] LCA antibody for B5 tissue

From:Patti Loykasek

    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> 
> Dear Histonetters,
> Do any of you know of a clone for CD45-LCA that stains B5 fixed
> tissue?  We are using a clone from DakoCytomation, anti-CD45, T29/33.  It
> stains formalin fixed tissue, but I see no staining in the B5 fixed tonsil
> control.  I have tried different blocks of tonsil with the same result.  All
> of the blocks stain with CD3 and CD20, so my guess is that this antibody
> does not recognize the antigen after B-5 fixation.
> Please let me know if you have had any success with CD45-LCA in B5
> fixed tissue.  Replies telling me not to use B5 are not helpful. I know it
> has a lot of negative qualities, and I would love to avoid the whole issue.
> Unfortunately, I have to work with an archive of tissue fixed in B5.
> Thanks for your assistance,
> Amanda Moklebust, HTL(QIHC)
> Seattle Cancer Care Alliance
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