Hylauronidase will digest the hyaluronic acid region of the molecule which
will unmask the access to BMP antigens.  There are three regions of the
molecule which we digest for, chondroitin sulfate (chondroitinase), keratin
sulfate (keratinase), and hyaluronic acid (hyaluronidase).  This is a
different concept than unwinding proteins which have been crosslinked by
aldehydes, in this case the actual physical structure of the molecule is
blocking the antigen site.  With some of these digestions you must also
include proteinase inhibitors to prevent loss of proteoglycans which may
what you are interested in marking.  We use proteinase inhibitors with
chondroitinase ABC digestion when we are doing IHC for Aggrecan, for
instance.
Patsy Ruegg

		-----Original Message-----
		From:	Renton, Lousie, Mrs
[mailto:177louie@chiron.wits.ac.za]
		Sent:	Tuesday, June 26, 2001 6:04 AM
		To:	histonet@pathology.swmed.edu
		Subject:	Haluronidase

		 Dear all

		A colleague of mine has an IHC method for bone morphogenic 
		proteins (on FFPE tissues) that uses an incubation in 
		mercapotoethanol and dithiothreitol followed by a
hyaluronidase 
		digestion. Could somebody please give an ex[planation for
these 
		steps?


		Thank you
		Louise Renton
		MRC
		Johannesburg