| Assay Method Information | |
| | Inhibitory Activity Assay |
| Description: | Table 2-3: Step 1. Add 50 μl anti-DIAPH1 (1:160 dilution in 0.1M NaHCO3 pH 9.6)/well. Incubate overnight at 4° C. Day 2 Step 2. Use the plate washer to aspirate anti-DIAPH11 and wash plates 4× with PBS 100 μl per well per wash. Step 3. Add 180 μl 3% BSA in 1×PBS. Incubate for 1.5 hrs at room temperature Step 4. Use the plate washer to aspirate the blocking solution and wash 5× with PBS 300 μl per well. Step 5. Add 50 μl of DIAPH1 containing lysate (85 μg) and incubate at RT for 3 hours. Step 6. Aspirate the lysate and wash the plate 5×(100 μl) on the plate washer. Step 7. Add 25 μl PBS to the wells. Step 8. Add 0.5 μL compound per well. Step 9. Add 24.5 μL GFP RAGE tail (125 nM) into each well for 2 hrs at room temperature. Step 10. Aspirate and wash 5× with PBS (100 μL) on the plate washer. Step 11. Add 100 μl PBS in each well. Step 12. Detection: Read on fluorescence plate reader excitation 435 nm and 485 nm emission |
| Affinity data for this assay | |
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