Assay Report Card

Basic Information

ID: ALA3705622

Type: Binding

Description: Inhibition Assay: The L-citrulline assay was based upon an original test-tube method developed by Prescott and Jones in 1969 (Prescott, L. M. & Jones, M. E. Modified methods for the determination of carbamyl aspartate. Anal Biochem 32, 408-419 (1969)), which was adapted and optimized for a microplate format. Subsequently, the activity of DDAH was quantified by detecting its conversion of ADMA to citrulline using the optimized protocol. The assay was scaled up to a 384-well format for high throughput chemical screening.High Throughput Screening of Small Molecules:Over 130,000 small molecules deposited in the Stanford High-throughput Bioscience Center (HTBC) were screened using the enzymatic assay to identify chemicals that regulate DDAH activity. In brief, recombinant human DDAH1 (rhDDAH1) was mixed with ADMA in the presence of screening buffer in 384-well plates using a Staccato multidrop. Small molecules (100 nL each) were then added to the wells using a robotic arrayer to yield a final compound screening.

Organism: Homo sapiens

Activity Charts

Compound Summary

Parent Molecular WeightALogPPolar Surface Area
151.191.591.59
217.180.810.81
230.263.273.27
401.492.452.45
293.391.991.99
322.413.353.35
213.201.821.82
204.230.980.98
203.272.232.23
341.442.952.95
347.483.023.02
393.583.653.65
323.421.621.62
369.493.213.21
371.422.122.12
375.883.093.09
385.492.832.83
345.473.083.08
295.360.840.84
361.853.043.04