Bookmark and Share
BioAssay: AID 651741

qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathway

Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration. The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of oxidative stress. The CellSensor ARE-bla HepG2 cell line (Invitrogen) can be used for analyzing the Nrf2/antioxidant response signaling pathway. Nrf2 (NF-E2-related factor more ..
_
   
 Tested Compounds
 Tested Compounds
All(2285)
 
 
Active(498)
 
 
Inactive(965)
 
 
Inconclusive(1030)
 
 
 Tested Substances
 Tested Substances
All(2810)
 
 
Active(583)
 
 
Inactive(1073)
 
 
Inconclusive(1154)
 
 
 Related BioAssays
 Related BioAssays
AID: 651741
Data Source: NCGC (ARE408)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-01

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 498
Related Experiments
AIDNameTypeComment
720653qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathway measured by Nrf2-dependant transcriptional activity - cell viability counter screenConfirmatorydepositor-specified cross reference
Description:
National Center for Advancing Translational Sciences [NCATS]
NIH Chemical Genomics Center [NCGC]
U.S. Environmental Protection Agency [EPA]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]

Assay Overview:

Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration. The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of oxidative stress. The CellSensor ARE-bla HepG2 cell line (Invitrogen) can be used for analyzing the Nrf2/antioxidant response signaling pathway. Nrf2 (NF-E2-related factor 2) and Nrf1 are transcription factors that bind to AREs and activate these genes. The CellSensor ARE-bla Hep G2 cell line contains a beta-lactamase reporter gene under control of the Antioxidant Response Element (ARE) stably integrated into HepG2 cells. The activation of the report gene under culture conditions can be detected by fluorescence intensity. This cell line can be used to screen chemicals in a 1536-well plate format and to identify agonists that induce oxidative stress.
Protocol
Assay Protocol Summary:
The ARE-bla cells were dispensed at 2,000 cells/5 ul/well in 1536-well black wall/clear bottom plates using a Multidrop dispenser. After the assay plates were incubated at a 37 C/5% CO2 incubator for 5 hours, 23 nL of compounds dissolved in DMSO, positive controls or DMSO only was transferred to the assay plate by a pin tool. The plates were incubated at 37 C for 16 hours. After 1 uL of LiveBLAzer B/G FRET substrate was added using a Flying Reagent Dispenser, the plates were incubated at room temperature for 2 hours, and fluorescence intensity was measured by an Envision plate reader.
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 activators are ranked higher than compounds that showed apparent inhibition.
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.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: HEPG2
From ChEMBL:
Assay Type: Functional
Result Definitions
Show more
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
5W460-Activity_ScoreActivity score.Integer
6W460-Curve_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 significant, but below 80% of control.String
7W460-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8W460-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9W460-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10W460-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11W460-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12W460-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13W460-Excluded_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
14W460-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15W460-Activity at 0.0008343153 uM (0.000834315μM**)% Activity at given concentration.Float%
16W460-Activity at 0.00279 uM (0.0027897μM**)% Activity at given concentration.Float%
17W460-Activity at 0.00590 uM (0.005901μM**)% Activity at given concentration.Float%
18W460-Activity at 0.014 uM (0.0139482μM**)% Activity at given concentration.Float%
19W460-Activity at 0.031 uM (0.0306175μM**)% Activity at given concentration.Float%
20W460-Activity at 0.068 uM (0.0684614μM**)% Activity at given concentration.Float%
21W460-Activity at 0.153 uM (0.153086μM**)% Activity at given concentration.Float%
22W460-Activity at 0.342 uM (0.342322μM**)% Activity at given concentration.Float%
23W460-Activity at 0.782 uM (0.781581μM**)% Activity at given concentration.Float%
24W460-Activity at 1.747 uM (1.74731μM**)% Activity at given concentration.Float%
25W460-Activity at 3.984 uM (3.9841μM**)% Activity at given concentration.Float%
26W460-Activity at 8.244 uM (8.2445μM**)% Activity at given concentration.Float%
27W460-Activity at 13.07 uM (13.0651μM**)% Activity at given concentration.Float%
28W460-Activity at 20.70 uM (20.7048μM**)% Activity at given concentration.Float%
29W460-Activity at 43.68 uM (43.6836μM**)% Activity at given concentration.Float%
30W460-Activity at 95.96 uM (95.9557μM**)% Activity at given concentration.Float%
31W460-Activity at 218.9 uM (218.902μM**)% Activity at given concentration.Float%
32W460-Activity at 489.5 uM (489.512μM**)% Activity at given concentration.Float%
33W460-Activity at 1094.5 uM (1094.48μM**)% Activity at given concentration.Float%
34W460-Activity at 2304.0 uM (2304μM**)% Activity at given concentration.Float%
35W460-Activity at 5812.0 uM (5812μM**)% Activity at given concentration.Float%
36W460-Activity at 13000.0 uM (13000μM**)% Activity at given concentration.Float%
37W460-Activity at 29060.0 uM (29060μM**)% Activity at given concentration.Float%
38W460-Activity at 64980.0 uM (64980μM**)% Activity at given concentration.Float%
39Ratio-Activity_ScoreActivity score.Integer
40Ratio-Curve_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 significant, but below 80% of control.String
41Ratio-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
42Ratio-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
43Ratio-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
44Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
45Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
46Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
47Ratio-Excluded_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
48Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
49Ratio-Activity at 0.0008343153 uM (0.000834315μM**)% Activity at given concentration.Float%
50Ratio-Activity at 0.00279 uM (0.0027897μM**)% Activity at given concentration.Float%
51Ratio-Activity at 0.00590 uM (0.005901μM**)% Activity at given concentration.Float%
52Ratio-Activity at 0.014 uM (0.0139482μM**)% Activity at given concentration.Float%
53Ratio-Activity at 0.031 uM (0.0306175μM**)% Activity at given concentration.Float%
54Ratio-Activity at 0.068 uM (0.0684614μM**)% Activity at given concentration.Float%
55Ratio-Activity at 0.153 uM (0.153086μM**)% Activity at given concentration.Float%
56Ratio-Activity at 0.342 uM (0.342322μM**)% Activity at given concentration.Float%
57Ratio-Activity at 0.782 uM (0.781581μM**)% Activity at given concentration.Float%
58Ratio-Activity at 1.747 uM (1.74731μM**)% Activity at given concentration.Float%
59Ratio-Activity at 3.984 uM (3.9841μM**)% Activity at given concentration.Float%
60Ratio-Activity at 8.244 uM (8.2445μM**)% Activity at given concentration.Float%
61Ratio-Activity at 13.07 uM (13.0651μM**)% Activity at given concentration.Float%
62Ratio-Activity at 20.70 uM (20.7048μM**)% Activity at given concentration.Float%
63Ratio-Activity at 43.68 uM (43.6836μM**)% Activity at given concentration.Float%
64Ratio-Activity at 95.96 uM (95.9557μM**)% Activity at given concentration.Float%
65Ratio-Activity at 218.9 uM (218.902μM**)% Activity at given concentration.Float%
66Ratio-Activity at 489.5 uM (489.512μM**)% Activity at given concentration.Float%
67Ratio-Activity at 1094.5 uM (1094.48μM**)% Activity at given concentration.Float%
68Ratio-Activity at 2304.0 uM (2304μM**)% Activity at given concentration.Float%
69Ratio-Activity at 5812.0 uM (5812μM**)% Activity at given concentration.Float%
70Ratio-Activity at 13000.0 uM (13000μM**)% Activity at given concentration.Float%
71Ratio-Activity at 29060.0 uM (29060μM**)% Activity at given concentration.Float%
72Ratio-Activity at 64980.0 uM (64980μM**)% Activity at given concentration.Float%
73W530-Activity_ScoreActivity score.Integer
74W530-Curve_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 significant, but below 80% of control.String
75W530-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
76W530-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
77W530-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
78W530-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
79W530-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
80W530-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
81W530-Excluded_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
82W530-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
83W530-Activity at 0.0008343153 uM (0.000834315μM**)% Activity at given concentration.Float%
84W530-Activity at 0.00279 uM (0.0027897μM**)% Activity at given concentration.Float%
85W530-Activity at 0.00590 uM (0.005901μM**)% Activity at given concentration.Float%
86W530-Activity at 0.014 uM (0.0139482μM**)% Activity at given concentration.Float%
87W530-Activity at 0.031 uM (0.0306175μM**)% Activity at given concentration.Float%
88W530-Activity at 0.068 uM (0.0684614μM**)% Activity at given concentration.Float%
89W530-Activity at 0.153 uM (0.153086μM**)% Activity at given concentration.Float%
90W530-Activity at 0.342 uM (0.342322μM**)% Activity at given concentration.Float%
91W530-Activity at 0.782 uM (0.781581μM**)% Activity at given concentration.Float%
92W530-Activity at 1.747 uM (1.74731μM**)% Activity at given concentration.Float%
93W530-Activity at 3.984 uM (3.9841μM**)% Activity at given concentration.Float%
94W530-Activity at 8.244 uM (8.2445μM**)% Activity at given concentration.Float%
95W530-Activity at 13.07 uM (13.0651μM**)% Activity at given concentration.Float%
96W530-Activity at 20.70 uM (20.7048μM**)% Activity at given concentration.Float%
97W530-Activity at 43.68 uM (43.6836μM**)% Activity at given concentration.Float%
98W530-Activity at 95.96 uM (95.9557μM**)% Activity at given concentration.Float%
99W530-Activity at 218.9 uM (218.902μM**)% Activity at given concentration.Float%
100W530-Activity at 489.5 uM (489.512μM**)% Activity at given concentration.Float%
101W530-Activity at 1094.5 uM (1094.48μM**)% Activity at given concentration.Float%
102W530-Activity at 2304.0 uM (2304μM**)% Activity at given concentration.Float%
103W530-Activity at 5812.0 uM (5812μM**)% Activity at given concentration.Float%
104W530-Activity at 13000.0 uM (13000μM**)% Activity at given concentration.Float%
105W530-Activity at 29060.0 uM (29060μM**)% Activity at given concentration.Float%
106W530-Activity at 64980.0 uM (64980μM**)% Activity at given concentration.Float%
107Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
PageFrom: