IFNα Receptor Binding in HUT78 Variants
| Cell Lines . | Kd (×10−10 M) . | Receptor Number/Cell . |
|---|---|---|
| HUT78 S1 | 5.8 ± 1.5 | 7500 ± 2100 |
| HUT78 S2 | 5.5 ± 2.0 | 7760 ± 2300 |
| HUT78 R1 | 6.0 ± 2.3 | 8010 ± 2500 |
| HUT78 R2 | 6.2 ± 2.3 | 8190 ± 2400 |
| Cell Lines . | Kd (×10−10 M) . | Receptor Number/Cell . |
|---|---|---|
| HUT78 S1 | 5.8 ± 1.5 | 7500 ± 2100 |
| HUT78 S2 | 5.5 ± 2.0 | 7760 ± 2300 |
| HUT78 R1 | 6.0 ± 2.3 | 8010 ± 2500 |
| HUT78 R2 | 6.2 ± 2.3 | 8190 ± 2400 |
Recombinant IFNα-2a was labeled with [125I] and purified by chromatography. The cells (1.5 × 106) were incubated with 5 pmol/L of [125I] IFNα-2a in 1 mL of RPMI medium/1% bovine serum albumin (BSA), with increasing amounts of unlabeled IFNα-2a for 2 hours at 4°C. The mixture was then centrifuged in dibutyl phthalate oil briefly to pellet the cells and counted in a gamma counter. The levels of IFNα binding to the HUT78 variants were determined by Scatchard analysis.