| Solution Information | help | |
| Enzyme: | Broad substrate specificity ATP-binding cassette transporter ABCG2 | |
| inhibitor: | BDBM47375 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The assay was a high throughput, singleplex no-wash procedure conducted in 384-well format micro plates in a total volume of 15.1 microliters dispensed sequentially as follows: 1) JC-1 substrate (10 microliter/well); 2) test compound (0.1 microliter/well), 3) drug-resistant unlabeled IgMXP3 cells (ABCG2) (5 microliter/well). Final in-well concentration of cells was 3 million cells/ml. Nine point dilution series were made of the test compound and depending on the experimental day the concentration of the test compound in-well varied from 66 micromolar to 0.015 micromolar. The plate contents were mixed, the plate rotated end-over-end at 4 RPM and 25 degrees C for 10 minutes, and then cell samples were immediately analyzed. Sample analysis was conducted with the HyperCyt(R) high throughput flow cytometry platform [Kuckuck, et al., 2001; Ramirez, et al., 2003]. Approximately 2 microliter volumes from each well were collected at a rate of approximately 40 samples per minute. This resulte | |
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