Re: [Histonet] ADG & Dysferlin troubles

From:Dana Settembre

I use Novocastra's Dysferlin cat.# NCL-Hamlet on human frozen muscle
 After I let the sections air dry from 
1 hour to overnight. I use it at a 1:160 dilution.
The detection system that I use is DakoCytomation's  LSAB2 kit and
their DAB kit.
Works well.
Dana Settembre
University Hospital-UMSNJ
Newark, NJ

>>> Karen Weidenheim  7/13/2005 9:10:34 AM
Dear All, we have never been able to get the Novocastra dysferlin Ab
work on frozen sections using DAB on the Ventana.  We have tried
multiple permutations.  Would someone be kind enough to share their
exact method for this antibody on Frozen sections of Muscle?
This is very important for diagnosis of muscular dystrophies.
Thanks so much.

Karen M. Weidenheim, M.D.
Professor of Pathology, Clinical Neurology and Clinical Neurosurgery
Albert Einstein College of Medicine
Chief, Division of Neuropathology
Montefiore Medical Center
111 East 210th Street
Bronx, NY   10467 
(718) 920-4446
FAX (718) 653-3409
Beeper (917) 556 3696
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>>> Dana Settembre  7/13/2005 7:43:36 AM >>>
I use  Dysferlin from Novocastra on frozen sections, but not FITC,
regular IHC. If you're desperate we can talk.
I use it with a detection kit for mouse, any vendor will usually do
DAB as the chromogen. No special scope is needed.
Dana Settembre
University Hospital-UMDNJ
Newark, NJ

>>> "Lewis, Sarah"  7/12/2005 1:48:17 PM >>>
Hello histoneters,
    I could really use any procedural information anyone has on
Alpha-dystroglycan (antibody only available thru Upstate) and
(antibody from VectorLabs).
        I have been attempting the stains for 2months now with no
satisfying results. I have tried several different
procedures/fixatives/incubation times/dilutions/. I perform a battery
14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green
with no background staining (picture perfect) .....except these two
boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO

Sarah E. Lewis 
Childrens Research 
Center for Gene Therapy 
700 Childrens Dr Rm WA3112 
Columbus OH 43205 

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