HIER and type II collagen

From:mrl0627@mail.ecu.edu


Thanks, Histonetters!
Your responses regarding HIER compatability with ab against type II collagen were a great help.  We had been running this antibody with fair success without AR.  The other antibodies I named had all improved GREATLY with the cooker and Citrate buffer.
My colleague was trying to run several antibodies on the same slide containing serial sections, and I was hoping that the poor response of type II to the treatment was due to some other problem, since the other abs she uses need the treatment.  Oh well!
We will, however, certainly try the enzymatic AR procedures suggested on Type II alone.
Incidently, I am the first person in our lab to even research AR and you all have been a priceless resource.  My thesis advisor calls my new pressure cooker the "Golden Retriever"!
Maureen
-----Original Message-----
From: mrl0627@mail.ecu.edu [mailto:mrl0627@mail.ecu.edu]
Sent: 10 July 2003 19:46
To: histonet@pathology.swmed.edu
Subject: HIER and type II collagen



Hello all!

We have been using antibody against type II collagen in mouse tissue fixed in Dent's and paraffin embedded successfully but have started using HIER with pressure cooker and citrate buffer and this antibody doesn't seem to be working (no staining in areas of limb cartilage that have always stained before). All other antibodies seem to love the AR (sections on same slide, incubated with antibodies against neurofilament, against myosin, against versican, all lit up beautifully). 
Does anybody know of a particular problem with 11-116B3 or other antibodies against type II collagen?
As always, TIA
Maureen 

								
			



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