Table 1.
Antibodies Used in this Study in Combination with Staining Conditions
| Antigen | Antibody | Species | Subtype | Titer | Fluorochrome | Treatment | Control tissue |
|---|---|---|---|---|---|---|---|
| EED | M26 | Mouse | IgG1 | 1:4000 o/n | strepAPC | Citrate ABC/CARD | Tonsil |
| EZH2 | K358 | Rabbit | 1:400 o/n | GaRALEXA 488 | EDTA ABC/CARD | Tonsil | |
| HPC1 | K350 | Rabbit | 1:100 o/n | GaRALEXA 488 | EDTA ABC/CARD | Kidney | |
| HPC2 | K326 | Rabbit | 1:100 o/n | GaRALEXA 488 | EDTA ABC/CARD | Tonsil | |
| HPH1 | K344 | Rabbit | 1:100 o/n | GaRALEXA 488 | EDTA ABC/CARD | Tonsil | |
| RING1 | K320 | Rabbit | 1:100 o/n | GaRALEXA 488 | Citrate ABC/CARD | Tonsil | |
| BMI-1 | K333 | Rabbit | 1:2000 o/n | GaRALEXA 488 | Citrate ABC/CARD | Tonsil | |
| BMI-1 | 6C9 | Mouse | IgG2b | undiluted | Rhodamine/tyramine | Citrate ABC/CARD | Tonsil |
| Ki67 | Mib-1 | Mouse | IgG1 | strepAPC | Citrate ABC/CARD | Tonsil | |
| MEL-18 | Rabbit | 1:400 o/n | GaRALEXA 488 | Citrate ABC/CARD | Tonsil | ||
| YY1 | Mouse | IgG1 | 1:250 o/n | strepAPC | Citrate ABC/CARD | Testis | |
| CtBP | Rabbit | 1:4000 o/n | GaRALEXA 488 | Citrate ABC/CARD | Testis | ||
| P16 | Mouse | IgG2a | 1:500 | Rhodamine/tyramine | Citrate ABC/CARD | Cervix carcinoma |
Abbreviations: o/n, overnight at 4°C; citrate, antigen retrieval using 0.01M citrate; pH 6.0, 15 minute in the autoclave; EDTA, antigen retrieval using 10mM EDTA, pH 8.0, 15 minute in the autoclave; strepAPC, allophycocyanin-conjugated streptavidine (blue signal); GaRALEXA, ALEXA 488-conjugated goat-anti-rabbit (green signal); rhodamine/tyramine (red signal); ABC, horseradish-peroxidase-conjugated streptavidin-biotin-complex; CARD, catalyzed reporter deposition method. For details about isolation and characterization of PcG-specific antisera: see references in Materials and Methods section.