RE: neonate mouse bones

If you can cut your samples without decal that is what you should do, you
can always put the cut sections in EDTA just before IHC staining to ensure
all the calcium has been removed which will cause problems with IHC
staining.  I do chondrocyte markers all the time on rat tibia and embyro rat
tissue, if I decal I use formic acid, it is good for IHC (Immunocal from
Decal Chemical).
Patsy Ruegg

		-----Original Message-----
		From:	Karen Glyde []
		Sent:	Tuesday, February 20, 2001 1:04 PM
		Subject:	neonate mouse bones

		Hello all,  I need to do  IHC on neonatal mouse bone days
4,13 and 18.  The
		particular interest is in the  epiphyseal plate of the tibia
		(chondrocytes)  The info I have rouned up suggests acid
decalcification is
		not appropriate   when followed by histochemical techniques
		investigations but rather the use of citrate buffer (pH4.5)
after a 24 to
		48 hour fix in 80% cold alcohol.  OR is bone this young
still cartilaginous
		enough not to need decalcification?   Reactions?
Suggestions?  Has someone
		done this?

		Karen L.Glyde
		Electron Microscope Facility
		Life Sciences Consortium
		Pennsylvania State University


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