Table 1. Antibodies used for IHC and IF staining.
| Antigen; product name | Host | Isot. | Target | Source |
|---|---|---|---|---|
|
| ||||
| PAB597 | m | NA | JCV VP1 | |
| SV40 T Ag (v-300); sc-20800 | r | IgG | JCV T Ag | Santa Cruz Biotec., Santa Cruz, CA |
| MAP-2; HM-2; M4403 | m | IgG1 | Neurons | Sigma, St-Louis, MO |
| MAP-2; ab5622 | r | IgG | Neurons | Chemicon, Temecula, CA |
| MAP-2; LS-B290 | ch | IgY | Neurons | Lifespan Bioscience, Seattle, WA |
| GFAP ; M0761; 6F2 | m | IgG1 | Astrocytes | Dako, Carpinteria, CA |
| GFAP ; Z0334 | r | IgG | Astrocytes | Dako, Carpinteria, CA |
| GFAP; ab4674 | ch | IgY | Astrocytes | Abcam, Cambridge, MA |
| CNPase; C5922; 11-5B | m | IgG1 | Myelin/Oligo | Sigma, St-Louis, MO |
| CNP; HPA023266 | r | IgG | Myelin/Oligo | Sigma, St-Louis, MO |
| MOG; EP4281; ab109746 | r | IgG | Myelin/Oligo | Abcam, Cambridge, MA |
| MAG; ab89780 | m | IgG1 | Myelin/Oligo | Abcam, Cambridge, MA |
| CD8; 1A5; VP-C325 | m | IgG1 | CD8 | Vector laboratories, Burlingame, CA |
| CD3; CD3-12; OASA11552 | rat | IgG | CD3 | Aviva Systems Biology, San Diego, CA |
| PLP | m | PLP | Serotec, USA | |
| NF | m | NF | Dako, Denmark | |
| KiM1P | m | KiM1P | Dr. Radzun, Goettingen, Germany | |
Isot: isotype, m: mouse, ch; chicken, r: rabbit, oligo: oligodendrocytes
For double staining, primary antibodies from different species (mouse, rat, rabbit or chicken) were used with appropriately matched (species and isotypes) secondary Abs. These antibodies were conjugated either to alkaline phosphatase (AP) or horseradish peroxidase (HRP) for IHC or to Alexa Fluor 350, 488 & 568 (Invitrogen), according to the manufacturer's instructions for IF. Single and double IHC were performed with the Vectastain Elite ABC Kits (Vector, Burlingame, CA), according to the manufacturer's instructions. Negative controls, for IHC and IFA, included omission of the primary Abs, use of isotype matched controls and use of sections from individuals known to be free of JCV-infected cells.