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BioAssay: AID 624476

Cytotoxicity counterscreen for NFkB agonists and antagonists

Cytotoxicity caused by screening compounds is a major concern for cell-based assays. We quantified cell death using a luminescence-based assay (Cell-Titer Glo, Promega), which determines the amount of ATP generated by living cells. High concentrations of Tetraoctylammonium and DMSO were used as positive and negative control, respectively ..more
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 Tested Compounds
 Tested Compounds
All(459)
 
 
Active(108)
 
 
Inactive(239)
 
 
Inconclusive(112)
 
 
 Tested Substances
 Tested Substances
All(461)
 
 
Active(109)
 
 
Inactive(239)
 
 
Inconclusive(113)
 
 
 Related BioAssays
 Related BioAssays
AID: 624476
Data Source: NCGC (MIPE3-NFKB-cv)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-08-13
Hold-until Date: 2013-01-01
Modify Date: 2013-01-03

Data Table ( Complete ):           Active    All
BioActive Compounds: 108
Depositor Specified Assays
AIDNameTypeComment
651645Cell Proliferation Assay against the TMD8 Cell Lineconfirmatory
651696Cell Proliferation Assay against a hMSC Cell Lineconfirmatory
651646Cell Proliferation Assay against the HBL1 Cell Lineconfirmatory
651556qHTS Assays for the Identification of Compounds that Synergize with a BTK Inhibitor (Ibrutinib, PCI-32765)summary
Description:
Cytotoxicity caused by screening compounds is a major concern for cell-based assays. We quantified cell death using a luminescence-based assay (Cell-Titer Glo, Promega), which determines the amount of ATP generated by living cells. High concentrations of Tetraoctylammonium and DMSO were used as positive and negative control, respectively
Protocol
Cells were trypsinized, washed in assay medium (AM, OPTI MEM with 0.5% fetal bovine serum, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 10mM HEPES pH 7.3, 100U/ml penicillin, and 100 ug/ml streptomycin) and plated at 2,000 cells/5 uL/well into white solid-bottom 1536-well plates. After an overnight incubation at 37C and 5% CO2, 23 nL of compound were delivered by a pin tool, and plates were incubated at 37C for 48 hrs. 2.5 uL of CellTiter-Glo (Promega) was added, the plates were centrifuged at 1,000 rpms and incubated at room temperature for ~15 minutes, and luminescence was measured at by a ViewLux (Perkin Elmer).
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
5Activity_ScoreActivity score.Integer
6Curve_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
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_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
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0003926931 uM (0.000392693μM**)% Activity at given concentration.Float%
16Activity at 0.0007804147 uM (0.000780415μM**)% Activity at given concentration.Float%
17Activity at 0.00118 uM (0.00117806μM**)% Activity at given concentration.Float%
18Activity at 0.00232 uM (0.00231672μM**)% Activity at given concentration.Float%
19Activity at 0.00353 uM (0.00353418μM**)% Activity at given concentration.Float%
20Activity at 0.00468 uM (0.00468253μM**)% Activity at given concentration.Float%
21Activity at 0.00703 uM (0.00703465μM**)% Activity at given concentration.Float%
22Activity at 0.014 uM (0.0138793μM**)% Activity at given concentration.Float%
23Activity at 0.021 uM (0.021085μM**)% Activity at given concentration.Float%
24Activity at 0.041 uM (0.0408261μM**)% Activity at given concentration.Float%
25Activity at 0.063 uM (0.0632551μM**)% Activity at given concentration.Float%
26Activity at 0.122 uM (0.122478μM**)% Activity at given concentration.Float%
27Activity at 0.190 uM (0.189642μM**)% Activity at given concentration.Float%
28Activity at 0.286 uM (0.286268μM**)% Activity at given concentration.Float%
29Activity at 0.563 uM (0.562965μM**)% Activity at given concentration.Float%
30Activity at 0.859 uM (0.858804μM**)% Activity at given concentration.Float%
31Activity at 1.138 uM (1.13785μM**)% Activity at given concentration.Float%
32Activity at 1.709 uM (1.70942μM**)% Activity at given concentration.Float%
33Activity at 3.373 uM (3.37266μM**)% Activity at given concentration.Float%
34Activity at 5.124 uM (5.12366μM**)% Activity at given concentration.Float%
35Activity at 9.921 uM (9.92074μM**)% Activity at given concentration.Float%
36Activity at 15.37 uM (15.371μM**)% Activity at given concentration.Float%
37Activity at 29.76 uM (29.7622μM**)% Activity at given concentration.Float%
38Activity at 46.08 uM (46.0829μM**)% Activity at given concentration.Float%
39Activity at 92.17 uM (92.1659μM**)% Activity at given concentration.Float%
40Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH084681 U54

Data Table (Concise)
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