Mary Vaughn was asking about CD4 and CD8 working on paraffin-embedded human
tissue.  I am working with human lung samples which are formalin-fixed and
paraffin
embedded, using the NovaCastra antibodies (NCL-CD4-1F6 and NCL-CD8-4b11) with
not much problem.  I autoclave the sections for 7 minutes in 1mM EDTA ph8.0
and then
do an APAAP method using the primaries at 1/50 dilution of each.  I use a
AS-BI
phosphate/New Fuchsin substrate so I can clear in xylne/Hemo-De and mount with
Entellan.  The only problem I have been having is some of the tissue (from
another
institution) keeps falling off in the autoclave, but because of suggestions
made earlier
on the Histonet I am trying to coat the slides with Parlodion/Celloidin to
keep them on
and it seems to be working, just needs a little more tweaking.  If anyone
wants furhter
info please contact me.


Mark Elliott, PhD
Research Associate
UBC-Pulmonary Research Lab
St. Paul's Hospital
Vancouver, BC Canada V6Z 1Y6
604-631-5351 (FAX)