[Histonet] CD marker help [particularly CD68]

[Amos Brooks]

Hi,
    It sounds like there are a number of antibodies with similar problems
here if you are having troubles with CD68, F4/80 and CD11b. If you are
getting variable results it could be that the tissues are not fixed and
processed sufficiently. If they are not fixed sufficiently there will be
inconsistencies in labeling of the epitopes. Adipose tissues take
particularly long to fix. In the absence of further details about the
procedure, that is where I would assume the gremlin is hiding.

Amos


On Tue, Jan 10, 2012 at 1:00 PM,
<histonet-request <@t> lists.utsouthwestern.edu>wrote:

> Message: 10
> Date: Tue, 10 Jan 2012 16:21:54 +0000
> From: "Till, Renee" <TillRenee <@t> uams.edu>
> Subject: [Histonet] CD marker help [particularly CD68]
> To: "histonet <@t> lists.utsouthwestern.edu"
>        <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>        <C4C329374E4EAC498FF4E294C530C63E35FB7521 <@t> COMMAIL2.ad.uams.edu>
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> Hello! I have been trying to get CD68 (CD11b and F4/80 as well, though I
> am focusing on the CD68 for now)on some mouse adipose tissue and have been
> getting varying results. I don't have any experience working with CD marker
> or doing IHC on adipose tissue. Without getting into the whole long process
> I have had trying to optimize this antibody, I am hoping someone can answer
> a couple of questions for me.
> 1. Can CD marker, and CD68 in particular, be picky  as far as needing
> fresh cut FFPE slides? With everything I have tried, this is the only thing
> I can come up with for the inconsistencies I have experienced.
> 2. Positive control. I do not have one. Any suggestions? At least I do not
> have a positive control adipose block. I have tried mouse spleen, lung, and
> duo, and all have worked, but not consistently, and not in keeping with my
> adipose test slides. I can have beautiful stained spleen and duo, but my
> adipose slide will be completely brown. I believe I have about worked this
> issue out....as far as getting good staining on the fat and the control
> tissues, but then I encounter what led to my question #1.
>
>
> Renee' Fox, HT (ASCP)
>