EPO Equilibrium Binding Studies on Selected Clones of 32D Cells Containing Variant Forms of the EPO-R
Cell Line . | kd (pmol/L) . | Receptor No./Cell . | R2 Value . |
---|---|---|---|
32D.EPO-R:1-483 (wild-type) | 220 260 | 834 1280 | .95 .93 |
345 | 1650 | .95 | |
32D.EPO-R:1-411 | 267 | 1413 | .94 |
325 | 1256 | .93 | |
383 | 1554 | .95 | |
32D.EPO-R:1-373 | 224 | 1300 | .96 |
258 | 1158 | .93 | |
373 | 1447 | .94 | |
32D.EPO-R:1-321 | 285 | 675 | .94 |
382 | 1055 | .95 | |
315 | 1252 | .94 | |
32D.EPO-R:1-252 | 650 | 6550 | .95 |
576 | 6882 | .93 | |
555 | 5897 | .94 | |
32D.EPO-R:1-483(W282R) | 402 | 1718 | .95 |
32D.EPO-R:1-483(YF) | 209 | 734 | .93 |
Cell Line . | kd (pmol/L) . | Receptor No./Cell . | R2 Value . |
---|---|---|---|
32D.EPO-R:1-483 (wild-type) | 220 260 | 834 1280 | .95 .93 |
345 | 1650 | .95 | |
32D.EPO-R:1-411 | 267 | 1413 | .94 |
325 | 1256 | .93 | |
383 | 1554 | .95 | |
32D.EPO-R:1-373 | 224 | 1300 | .96 |
258 | 1158 | .93 | |
373 | 1447 | .94 | |
32D.EPO-R:1-321 | 285 | 675 | .94 |
382 | 1055 | .95 | |
315 | 1252 | .94 | |
32D.EPO-R:1-252 | 650 | 6550 | .95 |
576 | 6882 | .93 | |
555 | 5897 | .94 | |
32D.EPO-R:1-483(W282R) | 402 | 1718 | .95 |
32D.EPO-R:1-483(YF) | 209 | 734 | .93 |
EPO binding studies were performed as described in Materials and Methods. EPO binding curves were plotted and the saturation of all EPO-binding sites was achieved for all cell lines over the concentrations of EPO used. For each cell line, at least 2 experiments were performed. Dissociation constant (kd) and cell surface receptor numbers were then determined through Scatchard analyses. R2values for all plots were greater than .93.