RE: Alcian Blue w/ & w/o Hyaluronidase

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From:Alex Brown <> (by way of histonet)
To:histonet <>
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Hi Joseph,
	We use ovine hyaluronidase ( hyalase )BP ( supplied in ampoules
of 1500 i.u. ) dissolved in 2 ml of phosphate buffer pH 6.7  -  3 hrs at
37 deg C    and 1% Alcian Blue in 3% Acetic Acid  (  pH 2.5  I
think.........  !  it's supposed to be 2.5 but we never actually measure
it ).    For positive control tissue we use umbilical cord.
	It's not 100% specific on its own, as John  Kiernan rightly
says, and we do sometimes get weak staining of other acid mucins.
However there should be an obvious difference between treated and
non-treated sections which is a fair indicator that hyaluronic acid is
present in the tissue.
			Alex Brown
			Crosshouse Hospital
			Kilmarnock, Scotland.
Subject: Alcian Blue w/ & w/o Hyaluronidase
Date: 26 February 1999 17:21

We have consistently had difficulty with the Alcian Blue with and
Hyaluronidase Stain.  We are using a section of cartilege as our
control. We use a Phosphate buffer solution at pH 5.6 with testicular
hyaluronidase purchased from Sigma Chemical.  We use 25 mg of
hyaluronidase to 50 cc of buffer. We are using a 3% solution of Alcian
Blue ph 2.5.  Any help out there????

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