Immunohistology of peripheral lymphocytes

From:David Choi <dchoi@nimr.mrc.ac.uk>

I'm currently working on the histology of peripheral nerve xenografts and
their rejection in a rat model. I've been trying to identify lymphocytes on
frozen sections of peripheral nerve xenografts, to look at the rejection
process, but am not having much success in labelling the lymphocytes.

I have tried OX1 (CD45) antibodies (1:200), and OX52 from Serotec (1:200) as
primaries, after 4% paraformaldehyde fixation, followed by Vector ABC
secondaries kit and DAB-Nickel-GOD. But I'm not getting good labelling of
the lymphocytes both in my experimental and control (spleen, thymus)
sections, despite fiddling with concentrations and incubation times.

Does anyone have a reliable protocol for pan-lymphocyte staining that would
work on cryo-sections of rat material? Many thanks!

David Choi
National Institute for Medical Research, London UK




<< Previous Message | Next Message >>