| Solution Information | help | |
| Enzyme: | Nucleotide-binding oligomerization domain-containing protein 2 | |
| inhibitor: | BDBM54431 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1. Hek293-T KbLuc cell line obtained from the assay provider's laboratory. 2. BrightGlo (Promega) 3. MDP (muramyl dipeptide) NOD2 Dose Response (This assay was multiplexed with a cytotoxicity protocol described in AID 1848). Procedure Day 1 1. Harvest HEK-293-T NF-kB -Luc at 100% confluency 2. Add 1 uL/well NOD assay media with Multidrop 3. Spin down plates @ 1000 RPM for 1 min. 4. Add with Echo 50nl total volume 100% DMSO to plates. *10pt serial dilution with Dilution Factor 2, starting from 20uM or 5uM Final assay concentrations. 1% DMSO Final assay concentration in 5uL. 5. Make cells suspension at 1.25X for final seeding density of 6000 cells/well in 4uL well volume. 6. Add MDP to 1.25X cell suspension at 0.875ug/mL. 7. Seed 6000 cells/well in 4uL/well to full plate HEK-293-T NF-kB to Corning # 3727 wht, 1536, hi-prof, TC-treated plate. 8. Spin down plates @ 500 RPM for 5 min. 9. Lid Plates. Sandwich each set 4 of four plates between two | |
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