You might want to try acetone fixation rather than aldehyde and anti-rat
antibodies,  I use antibodies from PharMingen, CD4, CD8, CD19, and CD25
Patsy Ruegg

		-----Original Message-----
		From:	David Choi [mailto:dchoi@nimr.mrc.ac.uk]
		Sent:	Thursday, December 28, 2000 5:01 AM
		To:	Histonet
		Subject:	Immunohistology of peripheral lymphocytes

		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