I did it on 5 micron paraffin sections of cartilage tissue.
What sort of "not getting good results" are you getting.. ie. background? uneven
staining? etc.
At that thickness it sounds as though you want whole nerves?
send me your protocol please.

John T. Demirs B. A. HTL (ASCP)
Wellman Laboratories of Photomedicine
Massachusetts General Hospital
Photopathology Dept. Edward 214
15 Fruit St.
Boston Ma 02114
617-726-6984 phone
617-726-1206 fax

> -----Original Message-----
> From:	Mitchell (Jane A.) [SMTP:jmitchell@neurology.wisc.edu]
> Sent:	Wednesday, February 12, 2003 12:58 PM
> To:	Demirs, John T.
> Subject:	RE: anti-protein gene product 9.5
> 
> John:  What thickness of sections are you working with?  I am using this
> antibody on 20 micron frozen sections of skin and not getting good
> results  - any helpful hints?
> Jean Mitchell, BS, HT (ASCP)
> University of Wisconsin Hospital & Clinics
> Department of Neurology
> Madison, WI
> 
> -----Original Message-----
> From: Demirs, John T. [mailto:DEMIRS@HELIX.MGH.HARVARD.EDU] 
> Sent: Wednesday, February 12, 2003 11:02 AM
> To: 'Martha Ward'
> Cc: Histo Net (E-mail)
> Subject: RE: anti-protein gene product 9.5
> 
> 
> PGP 9.5 Protocol
> 
> DAY ONE
> 1. Deparaffinization / Rehydrations to dH2O
> 2. Place slides in dH2O in 95 degree C water bath if using paraffin
> embedded sections ( microwave can be used at 3x1 min) 
> 3  Cool the slides to room temperature.
> 4. Treat with 0.3% H2O2 in methanol for 30 min.
> 5. Rinse in PBS 2 x 5 min.
> 6. Block in locking solution (4% NGS, 0.5% milk, 0.3% triton x-100 in
> PBS) 7. Incubate the tissue in PGP 9.5 overnight at Room temperature
> 1:1200 (dilute in 1/2 strength blocking solution)
> 
> DAY TWO
> 8. Rinse PBS 2 x 5 min
> 9. Incubate in secondary (biotinylated antirabbit IgG) 60 min.
> 10. Rinse
> PBS 2 x 5 min. 11. incubate in ABC (avidin biotinylated complex) 1 hour
> *note ABC must be made 30 mins. prior to use. 
> 12. Rinse 2 x 5 min.
> 13. Incubate in DAB 30 sec-5 min. 
> 14. dH2O 2 x10 min.
> 15. Counterstain if desired.
> 16  Dehydrate and coverslip
> 
> John T. Demirs B. A. HTL (ASCP)
> Wellman Laboratories of Photomedicine
> Massachusetts General Hospital
> Photopathology Dept. Edward 214
> 15 Fruit St.
> Boston Ma 02114
> 617-726-6984 phone
> 617-726-1206 fax
> 
> > -----Original Message-----
> > From:	Martha Ward [SMTP:mward@wfubmc.edu]
> > Sent:	Tuesday, February 11, 2003 1:54 PM
> > To:	histonet@pathology.swmed.edu
> > Subject:	anti-protein gene product 9.5
> > 
> > A researcher here is interested in working with the antibody 
> > anti-protein gene product 9.5.  Has anyone had any experience with
> this particular
> > antibody and also where did you purchase it?   Any information would
> be
> > greatly appreciated.  Thanks in advance.
> >  
> > Martha Ward, MT (ASCP) QIHC
> > Wake Forest University Baptist Medical Center
> > Winston-Salem, NC 27157