[Histonet] hyaluronidase digestion for aggrecan staining

[Hermina Borgerink]

Liz,

Many years ago when I was still routinely doing immuno staining on bone
and articular cartilage, this is the formula for hyaluronidase digestion
I used:

Hyaluronidase

0.1mg/ml bovine testicular hyaluronidase

0.05M Tris-HCl pH 7.6
0.15M NaCl
0.1% CaCl2
deionized water


Incubate sections at room temperature for 30 minutes prior to primary
antibody incubation.

Hermina

 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, August 29, 2006 1:28 PM
To: 'Histonet'
Subject: [Histonet] hyaluronidase digestion for aggrecan staining

Hello All

 

We have been working on aggrecan immunostain on articular cartiage and
wanted to perform a hyaluronidase digestion prior.  I have searched
extensively and have come up with few methods, none of which is
complete.
Does anyone out there have a method that includes the molarity of the
buffer, the pH of the buffer and the amount of hyaluronidase used and
the length of time and temp, etc..  Any advice would be helpful.

 

Thanks in advance

 

Liz

 

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC

Manager

Premier Laboratory, LLC

P.O. Box 18592

Boulder, CO 80308

phone (303) 735-5001

fax (303) 735-3540

liz <@t> premierlab.com

www.premierlab.com

 

Ship to Address:

 

Premier Laboratory, LLC

University of Colorado at Boulder

MCDB, Room A3B40

Boulder, CO 80309

 

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