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.
> 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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