RE: anti-protein gene product 9.5
PGP 9.5 Protocol
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)
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
> -----Original Message-----
> From: Martha Ward [SMTP:firstname.lastname@example.org]
> Sent: Tuesday, February 11, 2003 1:54 PM
> To: email@example.com
> 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
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