Bonnie, 
     Here is my procedure for syndecan. I use the Shandon Coverplate racks, 
     If you use a humidified chamber, decrease the incubation times by 50% 
     and do the rinses 1X PBS  Hope it helps.
     
     Sincerely, 
     Rachel Stock HTL(ASCP)
     Organongenesis, Inc.
     150 Dan Road
     Canton, MA 02021
     (781) 575-0775
     __________________________________________________________
     Date: 11 Jul 2000 15:54:39 -0500
     From: "Bonnie P Whitaker" <Bonnie.P.Whitaker@uth.tmc.edu>
     Subject: help with Syndecan-1,  FA1, SDF1, NRG-1
     
     >One of the pathologists that I am working with wants to do a project
     >involving the following antibodies:
     >Syndecan-1
     >FA1 (fetal antigen1)
     >SDF1 (stromal cell-derived factor 1)
     >NRG-1 (neuregulins)
     
     Antigen Retrieval Solution: (2X) 4.2g Citrate in 500ml dH2O + 500ml 
     0.1M NaOH, pH 6.0
     For final mix: 1:1 with dH2O on day of experiment
     1. Warm slides briefly in a 60oC oven, deparaffinize and rehydrate in 
     an automated stainer.
     2. Place slides face-up in a small plastic box, and cover with 1 inch 
     of Antigen Retrieval Solution.  Place the box in 1 inch deep H2O in a 
     glass tray. Microwave on high for 4 minutes (no boiling!). Cool 1-5 
     minutes.
     3. Incubate in endogenous peroxidase blocker according to blocking 
     protocol. (4 drops Zymed PeroxoBlock per well, incubate 5 minutes.)
     4. Rinse with Shandon Cadenza buffer.
     5. Set up slides in the Coverplate rack with 400 ml each of Cadenza 
     buffer 
     6. Make 20% normal goat serum in Cadenza buffer. Apply normal goat 
     serum to each slide (approx. 100ml), and incubate for 45 minutes at 
     room temperature.
     7. Dilute primary antibody 1:500 in PBS.  Apply primary antibody to 
     each slide, and incubate slides for 1-1.5 hours at room temperature.
     (My syndecan antibody is from Pharmingen.)
     8. Rinse slides twice with 2ml/well of Cadenza buffer.
     9. Secondary antibody: Biotinylated goat-a-rabbit from Vector 
     Vectastain Elite Kit: Mix 10ml PBS + 2 drops horse serum, add 1drop 
     secondary antibody. Apply secondary antibody to each slide (approx. 
     100ml), and incubate slides for 30-45 minutes at room temperature. 
     (make ABC reagent now.)
     10. Rinse slides twice with 2ml/well of Cadenza buffer.
     11. Link -from Vectastain Elite ABC/Mouse IgG kit:  1drop "A" + 1drop 
     "B" +2.5ml buffer
      Apply ABC reagent to each slide (approx. 100ml), and incubate for 
     30-45 minutes at room temperature.
     12. Rinse slides 2x 5 minutes with PBS or with 2ml/well of Cadenza 
     buffer.
     13. Prepare Zymed DAB reagent, apply to each slide, and incubate for 
     15 minutes at room temperature. 
     14. Rinse with distilled water, counterstain for 3 minutes with 
     hematoxylin, rinse in dH2O, dip in blueing reagent, then rinse again 
     in dH2O. Remove slides from the Coverplate cassettes. Dehydrate in the 
     automated stainer and coverslip using Xylene Substitute Mountant.