RE: Immunohistology of peripheral lymphocytes

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 []
		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

		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,
		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

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