96-well microplate coated with FN at 10μg/ml in PBS |
↓ |
Blocking with 2% BSA in PBS |
↓ |
Incubation with monoclonal antibodies |
↓ |
Incubation with trypsinized normal rat kidney(NRK)cells suspended in serum free D’MEM(10,000 cells per well) |
↓ |
Washing with PBS |
↓ |
Staining residual cells with 0.25% Rose Bengal dye in PBS |
↓ |
Washing with PBS |
↓ |
Treatment with 50% ethanol in PBS |
↓ |
Estimation of the number of attached cells by A570 |