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BioAssay: AID 1379

Counterscreen for Luciferase (Kinase-Glo TM) Inhibition

Luciferase (Kinase-Glo, Promega Corporation) was assayed for its ability to generate light using ATP and luciferin as substrates. The ATP concentration in the assay (10 uM) was within the linear range of enzyme activity for the assay conditions used. ..more
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 Tested Compounds
 Tested Compounds
All(199063)
 
 
Active(565)
 
 
Inactive(197526)
 
 
Inconclusive(982)
 
 
 Tested Substances
 Tested Substances
All(201160)
 
 
Active(565)
 
 
Inactive(199609)
 
 
Inconclusive(986)
 
 
AID: 1379
Data Source: NCGC (LUCKG089)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2008-08-13
Modify Date: 2010-07-06

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 565
Depositor Specified Assays
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AIDNameTypeComment
1771qHTS Assay for Inhibitors of CDC-like Kinase 4 (ADP-Glo Assay)confirmatory
2559Confirmation Assay for Inhibitors of Leishmania Mexicana Pyruvate Kinase (LmPK)confirmatory
1868qHTS Assay for Inhibitors of Human Galactokinase (GALK)confirmatory
2263Confirmation Concentration-Response Assay for Inhibitors of Human Muscle isoform 2 Pyruvate Kinaseconfirmatory
2506Confirmatory Assay for Inhibitors of Human Galactokinase (GALK): CDP-Me assay counterscreen for probe SARconfirmatory
2561Confirmation Assay for Inhibitors of Leishmania Mexicana Pyruvate Kinase (LmPK): for Probe SARconfirmatory
1969Confirmaiton Assay for Inhibitors of CDC-like Kinase 4 (ADP-Glo Assay)confirmatory
1770qHTS Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)confirmatory
1983Confirmation Assay for Inhibitors of CDC-like Kinase 4 (ADP-FP Assay)confirmatory
2035Confirmatory Assay for Inhibitors of Human Galactokinase (GALK): Phenol-HRP redox assay counterscreenconfirmatory
485316qHTS Assay for Rab9 Promoter Activators: Summarysummary
1634qHTS Assay for Inhibitors of Human Muscle isoform 2 Pyruvate Kinaseconfirmatory
2017Confirmatory Assay for Inhibitors of Human Galactokinase (GALK): CDP-Me assay counterscreenconfirmatory
1721qHTS Assay for Inhibitors of Leishmania Mexicana Pyruvate Kinase (LmPK)confirmatory
2015Confirmation Assay for Inhibitors of Human Galactokinase (GALK)confirmatory
2265Secondary assay for Inhibitors of Human Pyruvate Kinase M2 isoform: FLuc Counterscreenconfirmatory
2499Confirmation Assay for Inhibitors of Human Galactokinase (GALK): probe SARconfirmatory
2502Confirmatory Assay for Inhibitors of Human Galactokinase (GALK): Phenol-HRP redox assay counterscreen for probe SARconfirmatory
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2confirmatory
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)confirmatory
Description:
NCGC Assay Overview:

NIH Molecular Libraries Screening Centers Network [MLSCN]
NIH Chemical Genomics Center [NCGC]
MLSCN Grant: None

Luciferase (Kinase-Glo, Promega Corporation) was assayed for its ability to generate light using ATP and luciferin as substrates. The ATP concentration in the assay (10 uM) was within the linear range of enzyme activity for the assay conditions used.
Protocol
NCGC Assay Protocol Summary:

Four uL of substrate mix (final concentration, 50 mM imidazole pH 7.2, 50 mM KCl, 7 mM MgCl2, 10 uM ATP, 0.01% Tween 20, 0.05% BSA) was dispensed into white 1536 well plates. Twenty-three nL of compound was delivered by a pin tool and 2 uL of luciferase detection mix (Kinase-Glo, Promega)) was added. Following 8-minute incubation at ambient temperature, luminescence was measured by a ViewLux (Perkin Elmer) at 5 seconds exposure/plate. Data were normalized to the controls for basal activity (DMSO only) and 100% inhibition (40 uM SIB 1757) and curve fitting was performed using Hill equation.
Comment
Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically signficant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.0004904215 uM (0.000490421μM**)% Activity at given concentration.Float%
15Activity at 0.0004908046 uM (0.000490805μM**)% Activity at given concentration.Float%
16Activity at 0.00110 uM (0.00109693μM**)% Activity at given concentration.Float%
17Activity at 0.00117 uM (0.00116926μM**)% Activity at given concentration.Float%
18Activity at 0.00234 uM (0.00233851μM**)% Activity at given concentration.Float%
19Activity at 0.00245 uM (0.00245287μM**)% Activity at given concentration.Float%
20Activity at 0.00256 uM (0.0025567μM**)% Activity at given concentration.Float%
21Activity at 0.00468 uM (0.00467702μM**)% Activity at given concentration.Float%
22Activity at 0.00548 uM (0.00548314μM**)% Activity at given concentration.Float%
23Activity at 0.00549 uM (0.00548506μM**)% Activity at given concentration.Float%
24Activity at 0.00935 uM (0.00935405μM**)% Activity at given concentration.Float%
25Activity at 0.012 uM (0.0122644μM**)% Activity at given concentration.Float%
26Activity at 0.013 uM (0.0127835μM**)% Activity at given concentration.Float%
27Activity at 0.019 uM (0.0187081μM**)% Activity at given concentration.Float%
28Activity at 0.027 uM (0.0274153μM**)% Activity at given concentration.Float%
29Activity at 0.037 uM (0.0374162μM**)% Activity at given concentration.Float%
30Activity at 0.061 uM (0.0613176μM**)% Activity at given concentration.Float%
31Activity at 0.064 uM (0.0639184μM**)% Activity at given concentration.Float%
32Activity at 0.075 uM (0.0748324μM**)% Activity at given concentration.Float%
33Activity at 0.137 uM (0.137106μM**)% Activity at given concentration.Float%
34Activity at 0.150 uM (0.149665μM**)% Activity at given concentration.Float%
35Activity at 0.299 uM (0.29933μM**)% Activity at given concentration.Float%
36Activity at 0.307 uM (0.306569μM**)% Activity at given concentration.Float%
37Activity at 0.320 uM (0.319591μM**)% Activity at given concentration.Float%
38Activity at 0.599 uM (0.598659μM**)% Activity at given concentration.Float%
39Activity at 0.685 uM (0.685489μM**)% Activity at given concentration.Float%
40Activity at 1.197 uM (1.19732μM**)% Activity at given concentration.Float%
41Activity at 1.533 uM (1.53275μM**)% Activity at given concentration.Float%
42Activity at 1.598 uM (1.59796μM**)% Activity at given concentration.Float%
43Activity at 2.395 uM (2.39464μM**)% Activity at given concentration.Float%
44Activity at 3.427 uM (3.42724μM**)% Activity at given concentration.Float%
45Activity at 4.789 uM (4.78927μM**)% Activity at given concentration.Float%
46Activity at 7.663 uM (7.6633μM**)% Activity at given concentration.Float%
47Activity at 7.990 uM (7.98978μM**)% Activity at given concentration.Float%
48Activity at 9.579 uM (9.57854μM**)% Activity at given concentration.Float%
49Activity at 17.13 uM (17.1346μM**)% Activity at given concentration.Float%
50Activity at 17.14 uM (17.1351μM**)% Activity at given concentration.Float%
51Activity at 19.16 uM (19.1571μM**)% Activity at given concentration.Float%
52Activity at 38.31 uM (38.3142μM**)% Activity at given concentration.Float%
53Activity at 39.95 uM (39.9489μM**)% Activity at given concentration.Float%
54Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
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