Hi Amos, hi Marylou

We are using CD10, clone 56C6, Novocastra NCL-CD10-270, working conc. is 0.6
ug Ig/ml (corresponds to approx. 1:50, depending on lot - ask them to
specify Ig content, they have the info!), 60 min., RT. Pretreatment in 100
mM Tris-5% urea, pH 9.5 in microwave (pretty standard, ask back if you want
details about microwaving). Detection is with gt-a-mo Ig/Biotin (Dako E0433,
1:300) followed by StrABC/AP (Dako K0391, 1:200), each for 45 min., RT.
Works reasonably well, though some overfixed cases are difficult to stain.
CD5: clone 4C7, Novocastra NCL-CD5-4C7, working conc. is 0.9 ug Ig/ml;
otherwise same as above. Works well for T cells. Be aware that mantle cell
lymphoma cells express CD5 to a much lower degree than do T cells.
Therefore - and because of differences in fixation - in some cases you are
able to demonstrate CD5 in a mantle cell lymphoma, in other cases you are
not.
Clone 4C7 worked better in our hands than did CD5/54/B4 or CD5/54/F6. Good
luck!

Andi Kappeler
Institute of Pathology, University of Bern, Switzerland