>Hi Histonetters!
>
>	Once again, I am in desperate need of your help. I am trying to
>stain CD4 cells in human liver tissue. This tissue has been fixed and
>paraffin embedded. I have found many CD4 antibodies out there but they are
>all for either Flow Cytometry and/or Frozen Tissue sections. I have found
>one from Novocastra that will work with Paraffin embedded tissue, however,
>Antigen retrieval is required and I would prefer an antibody that would
>not require this step. I would appreciate any help anyone could give me.
>Thanks.
>
>Nima Farsinejad
>Research Specialist
>Digestive Diseases
>Emory University
>Atlanta, Georgia

**********

Hi Nima,

When your liver tissue is formaldehyde fixed, I don't think you will make
it without heath induced antigen retrieval. Staining without HIER will be
possible only if you fix your tissue in a non-crosslinking fixative. I can
warmly recommend to test methacarn [methanol:acetic acid
(glacial):chlorform = 6:1:3]. After fixation, don't forget to skip the
formalin step in the tissue processor!

If you wish to reveal CD4+ cells, how about double staining with CD3,rabbit
in red (AEC) and CD8,mouse in blue (Fast Blue). "Real" CD8+ cells will be
double-stained (purple), single stained cells should be the CD4-ones. Using
this trick you do not have the trouble making a distinction between CD4+
monocytes/macrophages.

Good luck.
Chris