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BioAssay: AID 410

p450-cyp1a2

The P450 gene superfamily is involved in metabolism and the clearance of xenobiotics. This assay used human CYP1A2 to measure the demethylation of luciferin 6' methyl ether (Luciferin-ME; Promega-Glo) to luciferin. The luciferin is then measured by luminescence after the addition of a luciferase detection regeant. Luciferin-ME concentration in the assay was equal to its Km for CYP1A2. ..more
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 Tested Compounds
 Tested Compounds
All(8354)
 
 
Active(4176)
 
 
Inactive(3676)
 
 
Inconclusive(713)
 
 
 Tested Substances
 Tested Substances
All(9198)
 
 
Active(4446)
 
 
Inactive(4019)
 
 
Inconclusive(733)
 
 
 Related BioAssays
 Related BioAssays
AID: 410
Data Source: NCGC
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2006-05-05
Modify Date: 2007-10-03

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 4176
Depositor Specified Assays
AIDNameTypeComment
899qHTS Assay for Inhibitors and Substrates of Cytochrome P450 2C19confirmatory
885qHTS Assay for Activators of Cytochrome P450 3A4confirmatory
883qHTS Assay for Inhibitors and Substrates of Cytochrome P450 2C9confirmatory
884qHTS Assay for Inhibitors and Substrates of Cytochrome P450 3A4confirmatory
891qHTS Assay for Inhibitors and Substrates of Cytochrome P450 2D6confirmatory
Description:
NCGC Assay Overview:

The P450 gene superfamily is involved in metabolism and the clearance of xenobiotics. This assay used human CYP1A2 to measure the demethylation of luciferin 6' methyl ether (Luciferin-ME; Promega-Glo) to luciferin. The luciferin is then measured by luminescence after the addition of a luciferase detection regeant. Luciferin-ME concentration in the assay was equal to its Km for CYP1A2.
Protocol
NCGC Assay Protocol Summary:

Two uL of enzyme and substrate mix (final concentrations: 20nM CYP1A2, 100uM Luciferin-ME, 100mM KPO4) were dispensed into a white, solid bottom 1536-well microtiter plate. 23nL of compound was transferred by a pin tool. The plate was incubated at room temperature for 10 min. Two uL of an NADPH regeneration solution (Promega supplied reagent) was dispensed and the reaction was allowed to continue for 60 min at room temperature. Then the reaction was stopped by reconstituted detection reagent. After another 20 min of incubation the plate was read on the ViewLux (Perkin Elmer) at 60 sec exposure and 2x2 binning. This protocol was carried out with Promega-Glo reagents and further information can be found at http://www.promega.com/tbs/tb325/tb325.pdf. Data were normalized to the to AC100 inhibition (57 uM furafylline).
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Activity DirectionIndicates direction of activity: inactive, decreasing, increasing.String
2Activity QualifierIndicates if AC50 is "=" to value or if it is infinite activity (i.e, inactive), ">".String
3Qualified AC50Qualified AC50; can be an IC50 or EC50.Float
4Log of AC50Log of AC50.Float
5Hill CoefficientHill slope of fitted curve.Float
6Curve R2R^2 fit value of curve. Closer to 1.0 equates to better Hill equation fit.Float
7Data TypeNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String
8Compound TypeNCGC designation for compound stage: 'NCGC Exploratory', 'NIHSMR', 'Compound Followup', 'Compound Verification', 'Probe Optimization'String
9Compound QCNCGC comment on compound QC: 'DPI QC', 'NCGC Purified'String
10Data Analysis QCNCGC comment on confidence of data: 'Curve Verfied', etcString
11NCGC CommentAnnotation/notes on a particular compound's data: e.g, 'cytotoxic', 'fluorescent artifact'String
12Curve Fit ModelCurve fitting model used.String
13Hill S0S0 of Hill EquationFloat
14Hill SinfSinf of Hill EquationFloat
15Hill dSdS of Hill EquationFloat
16Log AC50 Std ErrorStandard error for Log AC50.Float
17Curve Chi2fSqrt(X^2/f) of curve where X^2 is chi-squared. Lower number is a better fit.Float
18Excluded PointsIndividual points excluded from curve fitting.String
19Number of PointsNumber of valid points used for fitting.Integer
20Activity at 0.732nM% Activity at given concentration.Float%
21Activity at 1.636nM% Activity at given concentration.Float%
22Activity at 3.658nM% Activity at given concentration.Float%
23Activity at 8.18nM% Activity at given concentration.Float%
24Activity at 18.291nM% Activity at given concentration.Float%
25Activity at 0.041uM% Activity at given concentration.Float%
26Activity at 0.091uM% Activity at given concentration.Float%
27Activity at 0.204uM% Activity at given concentration.Float%
28Activity at 0.457uM% Activity at given concentration.Float%
29Activity at 1.022uM% Activity at given concentration.Float%
30Activity at 2.286uM% Activity at given concentration.Float%
31Activity at 5.111uM% Activity at given concentration.Float%
32Activity at 11.429uM% Activity at given concentration.Float%
33Activity at 25.556uM% Activity at given concentration.Float%
34Activity at 0.057mM% Activity at given concentration.Float%

Data Table (Concise)
Classification
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