[Histonet] Re: ADG & Dysferlin troubles

From:Jennifer Hofecker

Hi Everyone, sorry for the delay in responding, my
histonet digest was "lost in space" for a few days. 
We don't do ADG here, but I also use the Dysferlin
antibody from Novacastra (purchased from vector) with
regular IHC (no FITC) for frozen sections. I use Clone
Ham1/7B6(new vector number VP-D503).  
  Air dry frozen sections (usually 20 min or more). 
Dilution:  1:20 at rm temp 1 hr.
 I use with Vector R.T.U. vectastain kit (use
secondary 30 min@37degrees C. ABC reagent 1hr. rm
temp.  I use DAB from Vector for 15 min.  I usually do
a very pale counterstain or none at all depending on
I hope this is of some help to you.
Have a good weekend,

Jennifer Hofecker

Date: Wed, 13 Jul 2005 07:43:36 -0400
From: Dana Settembre 
Subject: Re: [Histonet] ADG & Dysferlin troubles
To: LewisS@ccri.net, histonet@lists.utsouthwestern.edu
Content-Type: text/plain; charset=US-ASCII

I use  Dysferlin from Novocastra on frozen sections,
but not FITC, just
regular IHC. If you're desperate we can talk.
I use it with a detection kit for mouse, any vendor
will usually do and
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 Dysferlin
(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!!!

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


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