RE: IHC on frozen muscle

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From:Sue Danielson <>
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Hello Louise,

We routinely perform dystrophin and sarcoglycan IHC on frozen muscle.  I
have not had any problems with sections sticking, even those Duchenne cases
which can be heavily infiltrated with fat & connective tissue.

We use poly-l-lysine coated slides (newcomber supply).  The muscle is
sectioned at a thickness of 7 microns, and slides are air-dried for a
minimum of 10 minutes.  We then begin staining with a 10 minute cold acetone
fixation in the refirgerator.  Slides are then pulled out of acetone, air
dried again and we proceed with our serum blocking step.  The rest of the
procedure is carried out on the bench, at room tmp.  The acetone step has
never given us any problems.  We have visualized the dystrophin with DAB,
Nova Red and also FITC in the past.
Hope this helps!

Susan Danielson, MS
Neuromuscular Lab Coordinator
Dept. Neurology, Medical College of Wisconsin
ph: 414.259.3836

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