| Assay Method Information | |
| | Tritiated Compound Binding to Membranes Isolated from Cells that Heterologous Express hNav1.7 and the beta1 Subunit |
| Description: | Preparation of membranes containing recombinantly expressed sodium channels: Following the procedure described in Example 243A, and making modifications as required to: centrifuge supernatants at 100000×g instead of 10000×g; re-suspend pellets in a buffer containing 0.01% BSA in addition to 100 mM NaCl, 20 mM Tris HCl, pH 7.4 buffer, and 1% v/v proteasome inhibitors; and storing membranes at −80° C. in single use aliquots instead of pooled.Radioligand Binding Studies: Saturation experiments: Following the procedure described in Example 243A, and making modifications as required to incubate [3H]compound for 3 hours instead of 18 hours.Competitive binding experiments: Following the procedure described in Example 243A, and making modifications as required to: perform binding experiments at room temperature for 3 hours instead of 18 hours; use 240 uL of solution and 300 pM of [3H]compound instead of 360 uL and 100 pM of [3H]compound; and wash filtered reactions 2 times instead of 5 times.Data Analysis: Following the procedure described in Example 243A. |
| Affinity data for this assay | |
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