Inhibition Assay: To determine their in vitro action on human PDE 5, the test substances are dissolved in 100% DMSO and serially diluted. Typically, d...

ID: ALA3706035

Type: Binding

Description: Inhibition Assay: To determine their in vitro action on human PDE 5, the test substances are dissolved in 100% DMSO and serially diluted. Typically, dilution series (1:3) from 200 uM to 0.091 uM are prepared (resulting final concentrations in the test: 4 uM to 0.0018 uM). In each case 2 ul of the diluted substance solutions are placed into the wells of microtitre plates (Isoplate-96/200 W; Perkin Elmer). Subsequently, 50 ul of a dilution of the above-described PDE 5 preparation are added. The dilution of the PDE 5 preparation is selected such that, during the later incubation, less than 70% of the substrate is converted (typical dilution: 1:100; dilution buffer: 50 mM tris/hydrochloric acid pH 7.5, 8.3 mM magnesium chloride, 1.7 mM EDTA, 0.2% BSA). The substrate, [8-3H]cyclic guanosine-3',5'-monophosphate (1 uCi/ul; Perkin Elmer) is diluted 1:2000 with assay buffer (50 mM tris/hydrochloric acid pH 7.5, 8.3 mM magnesium chloride, 1.7 mM EDTA).

Format: BAO_0000357

Organism: Homo sapiens

Target:  Phosphodiesterase 5A(ALA1827)

Document:  ALA3639285

BindingDB Source:  6895