| Solution Information | help | |
| Enzyme: | Integrin alpha-4 | |
| inhibitor: | BDBM75440 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The dose response assay is performed in 384-well micro titer plates in a total of 15 microL. Briefly, as with the primary HTS screen, the order of addition is: 5 microL of RPMI, followed by 0.1 microL of compound in DMSO at a series of concentrations (described below), followed by 5 microL of cell suspension, finishing with 5 microL of 36 nM LDV ligand with HUTS-21 PE antibodies. The negative control wells (low fluorescence) do not have any LDV, while the positive control wells (high fluorescence) do not have any compound. One compound is tested in 12 wells: negative control, positive control, nine compound concentrations from 3.1 nanoM to 20 microM, no cells (for time bin registration of the assay plate data file). Five microL of RPMI is added to the assay plate. Test compounds in DMSO are added and mixed (0.1 microL of a serial dilution, see below). Compounds are added to columns 3-11 and 15-23. Columns 1 and 13 are used for the negative control (no VLA-4 ligand added). Columns | |
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