Bookmark and Share
BioAssay: AID 2680

Counterscreen for JMJD2E Inhibitors: qHTS Assay for Inhibitors of Formaldehyde Dehydrogenase (FDH)

This assay is a counterscreen for a related assay for inhibitors of JMJD2E. See AID 2147 for details. The JMJD2E assay utilized an FDH-coupled protocol. ..more
_
   
 Tested Compounds
 Tested Compounds
All(137)
 
 
Active(16)
 
 
Inactive(114)
 
 
Inconclusive(7)
 
 
 Tested Substances
 Tested Substances
All(140)
 
 
Active(16)
 
 
Inactive(117)
 
 
Inconclusive(7)
 
 
AID: 2680
Data Source: NCGC (FDH2301)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-03-25

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 16
Related Experiments
AIDNameTypeComment
2147qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)Confirmatorydepositor-specified cross reference: qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)
2421Probe Summary for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)Summarydepositor-specified cross reference: Probe Summary for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)
2677Confirmation qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)Confirmatorysame project related to Summary assay
2687AlphaScreen Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)Confirmatorysame project related to Summary assay
2688MALDI-TOF-MS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)Confirmatorysame project related to Summary assay
493212Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E): 8HQs - Round 1Confirmatorysame project related to Summary assay
687009qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E): Counterscreen with LSD1Confirmatorysame project related to Summary assay
Description:
This assay is a counterscreen for a related assay for inhibitors of JMJD2E. See AID 2147 for details. The JMJD2E assay utilized an FDH-coupled protocol.

To screen for false positive results in the primary assay due to inhibition of the formaldehyde dehydrogenase coupled enzyme, an FDH-only assay was run using FDH as the enzyme (0.000025 U/uL final concentration) and 10 uM formaldehyde and 0.25 mM NAD+ as the substrate, and following the same fluidic protocol as described in AID 2147 for the coupled HTS assay.

Actives in this assay are likely false positives of the primary HTS.
Protocol
Assay is initiated by dispensing 3 uL of enzyme solution (4/3x final concentration) into a 1,536-well assay plate using a nanoliter solenoid-based dispenser (Beckman-Coulter, Fullerton, CA). After pintool transfer of compound solution, the assay plate is incubated at room temperature for 10 minutes, followed by a 1 uL addition of 4x substrate solution to initiate the reaction (final assay volume of 4 uL). The plate is then manually transferred into the ViewLux reader for kinetic measurements.
Comment
Keywords: FDH, JMJD2, JMJD2E, Human 2-Oxoglutarate Oxygenase, qHTS

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
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
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 significant, 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 0.0000096336 uM (9.63357e-06μM**)% Activity at given concentration.Float%
15Activity at 0.0000272478 uM (2.72478e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0000544957 uM (5.44957e-05μM**)% Activity at given concentration.Float%
17Activity at 0.0001089913 uM (0.000108991μM**)% Activity at given concentration.Float%
18Activity at 0.0002179827 uM (0.000217983μM**)% Activity at given concentration.Float%
19Activity at 0.0003758224 uM (0.000375822μM**)% Activity at given concentration.Float%
20Activity at 0.0009124032 uM (0.000912403μM**)% Activity at given concentration.Float%
21Activity at 0.00175 uM (0.00174986μM**)% Activity at given concentration.Float%
22Activity at 0.00319 uM (0.00318625μM**)% Activity at given concentration.Float%
23Activity at 0.00774 uM (0.00774329μM**)% Activity at given concentration.Float%
24Activity at 0.014 uM (0.0140534μM**)% Activity at given concentration.Float%
25Activity at 0.027 uM (0.0270158μM**)% Activity at given concentration.Float%
26Activity at 0.066 uM (0.0656545μM**)% Activity at given concentration.Float%
27Activity at 0.113 uM (0.112851μM**)% Activity at given concentration.Float%
28Activity at 0.229 uM (0.229117μM**)% Activity at given concentration.Float%
29Activity at 0.557 uM (0.557007μM**)% Activity at given concentration.Float%
30Activity at 0.907 uM (0.9073μM**)% Activity at given concentration.Float%
31Activity at 1.942 uM (1.94151μM**)% Activity at given concentration.Float%
32Activity at 5.423 uM (5.42267μM**)% Activity at given concentration.Float%
33Activity at 14.20 uM (14.1951μM**)% Activity at given concentration.Float%
34Activity at 23.41 uM (23.4054μM**)% Activity at given concentration.Float%
35Activity at 56.68 uM (56.6788μM**)% Activity at given concentration.Float%
36Compound QCSource of compound QCString

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

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