| Assay Method Information | |
| | Polarization Assay |
| Description: | eNOS was diluted to varying concentrations in buffer containing 50 mM inorganic phosphate, 50 mM KCl, and 5 mM DTT, pH 7.5. A spectrophotometer was used to determine the concentration of the fluorescein-labeled peptide using the extinction coefficient for fluorescein (ε492 nm = 79,000 M^-1). Peptides (100 nM) were added to samples containing varying concentrations of eNOS and incubated for 1 h at room temperature. Samples were then plated on 384-well Grenier Bio-One solid black microplates in triplicate. Polarization was measured with a Tecan Infinite M1000 Pro apparatus in fluorescent polarization mode using 470-nm excitation and 515-nm emission filters. |
| Affinity data for this assay | |
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