RE: merosin and emerin on frozen muscle sections.

From:"Houston, Ronnie"

In my experience, air-drying does not affect these particular antibody reactions. They do not require fixation either.
There are two merosin antibodies that are primarily used in neuromuscular diagnosis; the 80kD epitope from Chemicon, and the 300kD epitope from Novocastra. It is certainly true that the 80kD epitope has a more intense staining pattern than that of the 300kD.

Ronnie Houston
Regional Histology Operations Manager
Bon Secours HealthPartners Laboratories
5801 Bremo Road
Richmond, VA 23226
(804) 287 7972

-----Original Message-----
From: Tony Henwood []
Sent: Wednesday, June 04, 2003 6:45 PM
To: 'manal galal';
Subject: RE: merosin and emerin on frozen muscle sections.

Possibly air drying the sections is the problem for these particular antibodies. Have you tried alternatives? ethanol or 10% formalin or even acetone (though its fixative properties is questionable).

Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at  Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

-----Original Message-----
From: manal galal []
Sent: Wednesday, 4 June 2003 23:45
Subject: merosin and emerin on frozen muscle sections.

Dear colleagues,
             My work is on frozen muscle. I am having trouble with merosin and emerin immunostaining on frozen muscle. I wonder if my freezing process is to blame? 
    My freezing procedure is as follows:
1-We recieve the specimen immediately after the biopsy is taken.
2-We put it in a fridge for 15 min to relax.
3- The section is stuck on a pre-frozen chuck with a dab of OCT.
4- It is then immediately placed on the quick freeze shelf (-40C) of the cryostat for 15 min.
5-Then the specimen is put in the main compartment of the cryostat        (-20C) and left there for 20 min. to accomodate to the temperature.
6- The sections are then cut at 8 microns and placed on the slides.
7- The slides are left to dry overnight at room temperature.
By the way we get very good results with Dystrophins and Sarcoglycans, using this method of freezing.
Any suggestions??
Thank you
 Manal Galal

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