Bookmark and Share
BioAssay: AID 651772

qPCR Assay for CDK2 Expression Measured in Cell-Based System Using RT-PCR - 2084-11_Inhibitor_Dose_DryPowder_Activity

Assay Overview: MITF initiates a transcriptional program leading to melanocyte differentiation, cell cycle arrest, survival and pigmentation. It directly regulates the transcription of major cell cycle regulation genes including CDK2. SK-MEL-5 cells will be treated with compounds at dose. Cell extracts will be generated, mRNA will be purified and cDNA will be made from individual wells, qPCR will be performed for CDK2, an MITF target gene, and a house keeping gene, GAPDH. Compounds that suppress expression of MITF target gene at an AC50 < 10 uM will be pursued. ..more
_
   
 Tested Compounds
 Tested Compounds
All(34)
 
 
Active(29)
 
 
Inactive(1)
 
 
Inconclusive(4)
 
 
 Tested Substances
 Tested Substances
All(35)
 
 
Active(30)
 
 
Inactive(1)
 
 
Inconclusive(4)
 
 
AID: 651772
Data Source: Broad Institute (2084-11_Inhibitor_Dose_DryPowder_Activity)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-11-10
Hold-until Date: 2013-07-05
Modify Date: 2013-07-08

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 29
Related Experiments
Show more
AIDNameTypeProbeComment
488944Summary of Broad Institute MLPCN Identification of MITF Inhibitors ProjectSummary1 depositor-specified cross reference: Summary assay
488899MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_SinglePoint_HTS_ActivityScreening same project related to Summary assay
493073MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_DoseNoFile_CherryPick_Activity_Set3Confirmatory same project related to Summary assay
493102MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_DoseNoFile_CherryPick_Activity_Set4Confirmatory same project related to Summary assay
493177MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_DoseNoFile_CherryPick_Activity_Set2Confirmatory same project related to Summary assay
493191MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
493240MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_CherryPick_Activity_Set3Confirmatory same project related to Summary assay
540335MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_CherryPick_Activity_Set2Confirmatory same project related to Summary assay
540339MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
540346MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
540347MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
540348MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
624259MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatory same project related to Summary assay
624289MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
624290MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
624299MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
624315MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatory same project related to Summary assay
624316MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatory same project related to Summary assay
624324MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatory same project related to Summary assay
624362MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatory same project related to Summary assay
624363MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set5Confirmatory same project related to Summary assay
624364MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatory same project related to Summary assay
624366MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatory same project related to Summary assay
624368MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set5Confirmatory same project related to Summary assay
624426MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set7Confirmatory same project related to Summary assay
624427MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set5Confirmatory same project related to Summary assay
624428MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set7Confirmatory same project related to Summary assay
624429MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set6Confirmatory same project related to Summary assay
624430MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set8Confirmatory same project related to Summary assay
624440MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set6Confirmatory same project related to Summary assay
624488MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set6Confirmatory same project related to Summary assay
624489MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
624490MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set7Confirmatory same project related to Summary assay
624492MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set8Confirmatory same project related to Summary assay
651584MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_DryPowder_Activity_Set5Confirmatory same project related to Summary assay
651585MITF Orthogonal Assay: MALME3 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-04_Inhibitor_Dose_DryPowder_Activity_Set6Confirmatory same project related to Summary assay
651586MITF Orthogonal Assay: SK-MEL-5 CTG Assay Measured in Cell-Based System Using Plate Reader - 2084-02_Inhibitor_Dose_DryPowder_Activity_Set8Confirmatory same project related to Summary assay
651588MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set9Confirmatory same project related to Summary assay
651591MITF: Counter assay: A375 proliferation Measured in Cell-Based System Using Plate Reader - 2084-03_Inhibitor_Dose_DryPowder_Activity_Set9Confirmatory same project related to Summary assay
651753MITF Measured in Cell-Based System Using Plate Reader - 2084-01_Inhibitor_Dose_DryPowder_Activity_Set10Confirmatory same project related to Summary assay
651770qPCR Assay for TRPM1 Expression Measured in Cell-Based System Using RT-PCR - 2084-09_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651771qPCR Assay for DCT Expression Measured in Cell-Based System Using RT-PCR - 2084-12_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651773qPCR Assay for MITF Expression Measured in Cell-Based System Using RT-PCR - 2084-05_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651795qPCR Assay for MLANA Expression Measured in Cell-Based System Using RT-PCR - 2084-13_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651920Primary human melanocyte cytotoxicity assay Measured in Cell-Based System Using Plate Reader - 2084-06_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
Description:
Keywords: qPCR, MITF, SK-MEL-5, CDK2


Assay Overview: MITF initiates a transcriptional program leading to melanocyte differentiation, cell cycle arrest, survival and pigmentation. It directly regulates the transcription of major cell cycle regulation genes including CDK2. SK-MEL-5 cells will be treated with compounds at dose. Cell extracts will be generated, mRNA will be purified and cDNA will be made from individual wells, qPCR will be performed for CDK2, an MITF target gene, and a house keeping gene, GAPDH. Compounds that suppress expression of MITF target gene at an AC50 < 10 uM will be pursued.





Expected Outcome: Compounds that inhibit MITF directly or indirectly will reduce the expression of CDK2 and lead to an increased signal. Delta Ct of treated wells will be greater than delta Ct of mock wells; the Ct of the test gene is increased due to compounds, representing fewer mRNA copies. An increase in delta delta Ct is expected.
Protocol
Day 1 (Cell Plating)
SK-MEL-5 melanoma cells are added to each well of a Corning White, 384-well, tissue culture plate at 4,000 per well in 30 uL of DMEM media (10% FBS, 1% Pen/Strep/L-Glut) using a standard Combi liquid dispenser (Thermo). The plated cells are then incubated overnight in a tissue culture incubator (Thermo) at 37 degrees C and 5% CO2
Day 2 (Compound Pinning)
The following day, 100nL of compound is added to each well using the Cy-Bi Well pin transfer array. After addition of compound, the plates are returned to the tissue culture incubator and are again incubated overnight.
Day 3 (cDNA Synthesis)
1. Cell lysis (using Cells-to-Ct kits, Ambion cat.no. 4391851C)
The medium is aspirated and the cells are washed twice (100 uL with PBS) using the ELX405 Plate washer (Biotek). The assay plates are flipped upside down and centrifuged at 1000 rpm 2 minutes to remove the excess liquid. 10 uL of Lysis solution with DNase I (from Cell to CT Lysis kit, Ambion) is added to each well using the MultiDrop Combi/Standard tube dispensing cassette (Thermo Scientific). Each assay plate is then shaken for 2 minutes and incubated for an additional 5 minutes at room temperature. At the end of the incubation, 1 uL of stop solution (from Cells to CT Lysys kit, Ambion) is added with the Multidrop Combi-nl (Thermo Scientific) and the assay plates are centrifuged (faced up)(1000 rpm, 2minutes) . The assay plates are incubated for 2 minutes at room temperature and processed for reverse transcription.
2. Reverse transcription (RT) (using Cell to CT RT kit, Ambion cat. no. #4402958)
TABLE 1. 8 uL RT Master Mix/10 uL total reaction volume)
Component Amount per reaction
2X RT Buffer 5 uL
20X RT Enzyme Mix 0.5 uL
Nuclease-Free Water 2.5 uL
3. 8 uL of RT is dispensed into each well of a RT assay plate (Axygen, PCR-384 RGD C). 2 uL of the lysed cells are transferred into RT assay plate using Vario transfer unit (CyBi Well). The RT assay plates are incubated at 37 degrees C for 1h and the reverse transcriptase is inactivated by incubating the plates for 1 minute at 95 degrees C. cDNA is stored at -80 degrees C until ready for qPCR analysis.
Day 4 (qPCR)
1. Prepare qPCR master mix (using Light Cycler Probes Master, Roche cat. no.4887301001)
Table 2.
4 uL PCR Master Mix/5uL total volume of reaction
Component ###Amount per reaction
2X Roche Master Mix (Probes Master) ###2.5 uL
20X FAM Taqman probe/primers set ###0.125 uL
(human CDK2, Applied Biosystems, 4331182 Hs01548894_m1 )
20X VIC Taqman probe/primers set ###0.125 uL
(human GAPDH, Applied Biosystems C10228)PCR H2O ###1.25 uL
2. 4 uL/well of PCR master mix is dispensed in PCR plate (Roche Light Cycler 480 MultiWell Plate 384, Cat# 04 729 749 001) using the Multidrop Combi-nl (Thermo Scientific).
3. 1 uL/well of RT DNA is transferred in the 4 uL/well PCR plate. The PCR plates are centrifuged (faced up) 2 minutes at 1000 rpm.
4. PCR is performed using ThermoCycler (Roche Light Cycler 480 II) with Macro Probe Protocol with this setup:
Step Temperature Time
1. 95 degrees C 10 minutes
2. 95 degrees C 10 seconds
3. 60 degrees C 30 seconds
Step 2 and 3 (55 cycles)
4.#40 degrees C#### 30 seconds
Comment
1. Using the Roche Light Cycler instrument software, a cycle call was generated for when each well enters log phase amplification (Ct).
2. The delta Ct value was determined by subtracting the Ct value of the control gene (GAPDH) from the Ct value of the target gene (CDK2) in each well.
3. The delta delta Ct value of each compound treatment was determined by averaging the delta Ct values of the mock well on each plate and subtracting that average from the delta Ct value of each compound well.
PRESENCE OF CONTROLS: Neutral control wells (NC; n=36) were included on every plate.
EXPECTED OUTCOME: Active compounds result in increase readout signal.
ACTIVE CONCENTRATION LIMIT:
For each sample, the highest valid tested concentration (Max_Concentration) was determined and the active concentration limit (AC_limit) was set to equal (2)(Max_Concentration).
NORMALIZATION:
The raw signals of the plate wells were normalized using the 'Neutral Controls' method in Genedata Assay Analyzer (v.10.0.2):
The median raw signal of the intraplate neutral controls (NC) is set to a normalized activity value of 0.
A normalized activity value of 100 is defined as (2)(NC).
A normalized activity value of 50 is defined as (0.5)(NC).
Experimental wells values were scaled to this range.
PATTERN CORRECTION: No plate pattern correction algorithm from Genedata Condoseo (v.10.0.2) was applied.
MEASUREMENT USED TO DETERMINE ACTIVE CONCENTRATION (AC): AbsACNN where NN=40
AbsACNN values were calculated using the curve fitting strategies in Genedata Screener Condoseo (v.10.0.2).
AbsACNN values were calculated up to the active concentration limit described for each sample.
pAbsACNN was set to equal -1*log10(AbsACNN)
PUBCHEM_ACTIVITY_OUTCOME:
Activity_Outcome = 1 (inactive) when:
a) compound shows activity but in a direction opposite to the expected outcome
in these cases, values describing curve fitting parameters (Sinf, S0, Hill Slope, log_AbsACNN, log_AbsACNN_SE) are set to null
b) curve fit is constant where activity is > -30% and < 30% at all tested concentrations, or
c) AbsACNN> AbsACNN_limit
Activity_Outcome = 2 (active) when:
AbsACNN <= AbsACNN_limit
Activity_Outcome = 3 (inconclusive) when:
a) Curve fitting strategy resulted in a constant fit with activity >= -70% but <= -30%, or
b) The fit was deemed not valid due to poor fit quality.
PUBCHEM_ACTIVITY_SCORE:
If PUBCHEM_ACTIVITY_OUTCOME = 1 (inactive) or 3 (inconclusive),
then PUBCHEM_ACTIVITY_SCORE = 0
If PUBCHEM_ACTIVITY_OUTCOME = 2 (active)
then PUBCHEM_ACTIVITY_SCORE = (10)(pAbsACNN)
Scores relate to AbsACNN in this manner:
120 = 1 pM
90 = 1 nM
60 = 1 uM
30 = 1 mM
0 = 1 M
When the active concentration (AbsACNN) is calculated to be greater than the highest valid tested concentration (Max_Concentration), the PUBCHEM_ACTIVITY_SCORE is calculated using Max_Concentration as the basis.
When the active concentration (AbsACNN) is calculated to be less than the lowest tested concentration, the PUBCHEM_ACTIVITY_SCORE is calculated using the lowest tested concentration as the basis.
Note:
The individual dose data point columns ('Activity_at_xxuM') reported here represent the median of valid (unmasked) replicate observations at each concentration. These values are the inputs to a curve fitting algorithm.
All other data columns represent values which are derived during the curve fitting algorithm; this may sometimes include automatic further masking of some replicate data points.
Occasionally this results in perceived inconsistencies: for example, between the derived 'Maximal_Activity' and the apparent most active data point.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Cell-based
Assay Cell Type: SK-MEL-5
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1AbsAC40_Qualifier<, =, or >String
2AbsAC40_uM*The concentration at which the fitted curve passes activity threshold 40.FloatμM
3pAbsAC40_MEqual to -1*log10(AbsAC40).Float
4ACTIVITY_OUTCOME_TEST1The PubChem Activity Outcome for the subset of data included in the indicated Test number.Integer
5ACTIVITY_SCORE_TEST1The PubChem Activity Score for the subset of data included in the indicated Test number.Integer
6ASSAYDATA_COMMENT_TEST1Comments relevant to the subset of data included in the indicated Test number.String
7AbsAC40_Qualifier_TEST1<, =, or >String
8AbsAC40_uM_TEST1The concentration at which the fitted curve passes activity threshold nn.FloatμM
9pAbsAC40_M_TEST1Equal to -1*log10(AbsACnn).Float
10Hill_Slope_TEST1The slope at AC50Float
11S0_(delta-delta-Ct)_TEST1The fitted activity value at zero concentrationFloat%
12Sinf_(delta-delta-Ct)_TEST1The fitted activity value at infinite concentrationFloat%
13Num_Points_TEST1The number of data points used to generate the plotInteger
14Max_Activity_(delta-delta-Ct)_TEST1The maximum activity value observed, based on mean of replicates per concentrationFloat%
15Max_Activity_Conc_uM_TEST1The concentration at which the maximum activity is observedFloatμM
16Max_Concentration_uM_TEST1Maximum valid test concentrationFloatμM
17Activity_at_0.000060uM_(delta-delta-Ct)_TEST1 (6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
18Activity_at_0.00012uM_(delta-delta-Ct)_TEST1 (0.00012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
19Activity_at_0.00026uM_(delta-delta-Ct)_TEST1 (0.00026μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
20Activity_at_0.0005uM_(delta-delta-Ct)_TEST1 (0.0005μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
21Activity_at_0.001uM_(delta-delta-Ct)_TEST1 (0.001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
22Activity_at_0.0021uM_(delta-delta-Ct)_TEST1 (0.0021μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
23Activity_at_0.0042uM_(delta-delta-Ct)_TEST1 (0.0042μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
24Activity_at_0.008uM_(delta-delta-Ct)_TEST1 (0.008μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
25Activity_at_0.016uM_(delta-delta-Ct)_TEST1 (0.016μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
26Activity_at_0.03uM_(delta-delta-Ct)_TEST1 (0.03μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
27Activity_at_0.068uM_(delta-delta-Ct)_TEST1 (0.068μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
28Activity_at_0.135uM_(delta-delta-Ct)_TEST1 (0.135μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
29Activity_at_0.26uM_(delta-delta-Ct)_TEST1 (0.26μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
30Activity_at_0.5uM_(delta-delta-Ct)_TEST1 (0.5μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
31Activity_at_1uM_(delta-delta-Ct)_TEST1 (1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
32Activity_at_2.1uM_(delta-delta-Ct)_TEST1 (2.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
33Activity_at_4.2uM_(delta-delta-Ct)_TEST1 (4.2μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
34Activity_at_8uM_(delta-delta-Ct)_TEST1 (8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
35Activity_at_16uM_(delta-delta-Ct)_TEST1 (16μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
36Activity_at_35uM_(delta-delta-Ct)_TEST1 (35μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
37ACTIVITY_OUTCOME_TEST2The PubChem Activity Outcome for the subset of data included in the indicated Test number.Integer
38ACTIVITY_SCORE_TEST2The PubChem Activity Score for the subset of data included in the indicated Test number.Integer
39ASSAYDATA_COMMENT_TEST2Comments relevant to the subset of data included in the indicated Test number.String
40AbsAC40_Qualifier_TEST2<, =, or >String
41AbsAC40_uM_TEST2The concentration at which the fitted curve passes activity threshold nn.FloatμM
42pAbsAC40_M_TEST2Equal to -1*log10(AbsACnn).Float
43Hill_Slope_TEST2The slope at AC50Float
44S0_(delta-delta-Ct)_TEST2The fitted activity value at zero concentrationFloat%
45Sinf_(delta-delta-Ct)_TEST2The fitted activity value at infinite concentrationFloat%
46Num_Points_TEST2The number of data points used to generate the plotInteger
47Max_Activity_(delta-delta-Ct)_TEST2The maximum activity value observed, based on mean of replicates per concentrationFloat%
48Max_Activity_Conc_uM_TEST2The concentration at which the maximum activity is observedFloatμM
49Max_Concentration_uM_TEST2Maximum valid test concentrationFloatμM
50Activity_at_0.000060uM_(delta-delta-Ct)_TEST2 (6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
51Activity_at_0.00012uM_(delta-delta-Ct)_TEST2 (0.00012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
52Activity_at_0.00026uM_(delta-delta-Ct)_TEST2 (0.00026μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
53Activity_at_0.0005uM_(delta-delta-Ct)_TEST2 (0.0005μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
54Activity_at_0.001uM_(delta-delta-Ct)_TEST2 (0.001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
55Activity_at_0.0021uM_(delta-delta-Ct)_TEST2 (0.0021μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
56Activity_at_0.0042uM_(delta-delta-Ct)_TEST2 (0.0042μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
57Activity_at_0.008uM_(delta-delta-Ct)_TEST2 (0.008μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
58Activity_at_0.016uM_(delta-delta-Ct)_TEST2 (0.016μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
59Activity_at_0.03uM_(delta-delta-Ct)_TEST2 (0.03μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
60Activity_at_0.068uM_(delta-delta-Ct)_TEST2 (0.068μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
61Activity_at_0.135uM_(delta-delta-Ct)_TEST2 (0.135μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
62Activity_at_0.26uM_(delta-delta-Ct)_TEST2 (0.26μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
63Activity_at_0.5uM_(delta-delta-Ct)_TEST2 (0.5μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
64Activity_at_1uM_(delta-delta-Ct)_TEST2 (1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
65Activity_at_2.1uM_(delta-delta-Ct)_TEST2 (2.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
66Activity_at_4.2uM_(delta-delta-Ct)_TEST2 (4.2μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
67Activity_at_8uM_(delta-delta-Ct)_TEST2 (8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
68Activity_at_16uM_(delta-delta-Ct)_TEST2 (16μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
69Activity_at_35uM_(delta-delta-Ct)_TEST2 (35μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
70ACTIVITY_OUTCOME_TEST3The PubChem Activity Outcome for the subset of data included in the indicated Test number.Integer
71ACTIVITY_SCORE_TEST3The PubChem Activity Score for the subset of data included in the indicated Test number.Integer
72ASSAYDATA_COMMENT_TEST3Comments relevant to the subset of data included in the indicated Test number.String
73AbsAC40_Qualifier_TEST3<, =, or >String
74AbsAC40_uM_TEST3The concentration at which the fitted curve passes activity threshold nn.FloatμM
75pAbsAC40_M_TEST3Equal to -1*log10(AbsACnn).Float
76Hill_Slope_TEST3The slope at AC50Float
77S0_(delta-delta-Ct)_TEST3The fitted activity value at zero concentrationFloat%
78Sinf_(delta-delta-Ct)_TEST3The fitted activity value at infinite concentrationFloat%
79Num_Points_TEST3The number of data points used to generate the plotInteger
80Max_Activity_(delta-delta-Ct)_TEST3The maximum activity value observed, based on mean of replicates per concentrationFloat%
81Max_Activity_Conc_uM_TEST3The concentration at which the maximum activity is observedFloatμM
82Max_Concentration_uM_TEST3Maximum valid test concentrationFloatμM
83Activity_at_0.000060uM_(delta-delta-Ct)_TEST3 (6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
84Activity_at_0.00012uM_(delta-delta-Ct)_TEST3 (0.00012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
85Activity_at_0.00026uM_(delta-delta-Ct)_TEST3 (0.00026μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
86Activity_at_0.0005uM_(delta-delta-Ct)_TEST3 (0.0005μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
87Activity_at_0.001uM_(delta-delta-Ct)_TEST3 (0.001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
88Activity_at_0.0021uM_(delta-delta-Ct)_TEST3 (0.0021μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
89Activity_at_0.0042uM_(delta-delta-Ct)_TEST3 (0.0042μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
90Activity_at_0.008uM_(delta-delta-Ct)_TEST3 (0.008μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
91Activity_at_0.016uM_(delta-delta-Ct)_TEST3 (0.016μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
92Activity_at_0.03uM_(delta-delta-Ct)_TEST3 (0.03μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
93Activity_at_0.068uM_(delta-delta-Ct)_TEST3 (0.068μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
94Activity_at_0.135uM_(delta-delta-Ct)_TEST3 (0.135μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
95Activity_at_0.26uM_(delta-delta-Ct)_TEST3 (0.26μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
96Activity_at_0.5uM_(delta-delta-Ct)_TEST3 (0.5μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
97Activity_at_1uM_(delta-delta-Ct)_TEST3 (1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
98Activity_at_2.1uM_(delta-delta-Ct)_TEST3 (2.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
99Activity_at_4.2uM_(delta-delta-Ct)_TEST3 (4.2μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
100Activity_at_8uM_(delta-delta-Ct)_TEST3 (8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
101Activity_at_16uM_(delta-delta-Ct)_TEST3 (16μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
102Activity_at_35uM_(delta-delta-Ct)_TEST3 (35μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
103ACTIVITY_OUTCOME_TEST4The PubChem Activity Outcome for the subset of data included in the indicated Test number.Integer
104ACTIVITY_SCORE_TEST4The PubChem Activity Score for the subset of data included in the indicated Test number.Integer
105ASSAYDATA_COMMENT_TEST4Comments relevant to the subset of data included in the indicated Test number.String
106AbsAC40_Qualifier_TEST4<, =, or >String
107AbsAC40_uM_TEST4The concentration at which the fitted curve passes activity threshold nn.FloatμM
108pAbsAC40_M_TEST4Equal to -1*log10(AbsACnn).Float
109Hill_Slope_TEST4The slope at AC50Float
110S0_(delta-delta-Ct)_TEST4The fitted activity value at zero concentrationFloat%
111Sinf_(delta-delta-Ct)_TEST4The fitted activity value at infinite concentrationFloat%
112Num_Points_TEST4The number of data points used to generate the plotInteger
113Max_Activity_(delta-delta-Ct)_TEST4The maximum activity value observed, based on mean of replicates per concentrationFloat%
114Max_Activity_Conc_uM_TEST4The concentration at which the maximum activity is observedFloatμM
115Max_Concentration_uM_TEST4Maximum valid test concentrationFloatμM
116Activity_at_0.000060uM_(delta-delta-Ct)_TEST4 (6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
117Activity_at_0.00012uM_(delta-delta-Ct)_TEST4 (0.00012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
118Activity_at_0.00026uM_(delta-delta-Ct)_TEST4 (0.00026μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
119Activity_at_0.0005uM_(delta-delta-Ct)_TEST4 (0.0005μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
120Activity_at_0.001uM_(delta-delta-Ct)_TEST4 (0.001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
121Activity_at_0.0021uM_(delta-delta-Ct)_TEST4 (0.0021μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
122Activity_at_0.0042uM_(delta-delta-Ct)_TEST4 (0.0042μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
123Activity_at_0.008uM_(delta-delta-Ct)_TEST4 (0.008μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
124Activity_at_0.016uM_(delta-delta-Ct)_TEST4 (0.016μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
125Activity_at_0.03uM_(delta-delta-Ct)_TEST4 (0.03μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
126Activity_at_0.068uM_(delta-delta-Ct)_TEST4 (0.068μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
127Activity_at_0.135uM_(delta-delta-Ct)_TEST4 (0.135μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
128Activity_at_0.26uM_(delta-delta-Ct)_TEST4 (0.26μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
129Activity_at_0.5uM_(delta-delta-Ct)_TEST4 (0.5μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
130Activity_at_1uM_(delta-delta-Ct)_TEST4 (1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
131Activity_at_2.1uM_(delta-delta-Ct)_TEST4 (2.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
132Activity_at_4.2uM_(delta-delta-Ct)_TEST4 (4.2μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
133Activity_at_8uM_(delta-delta-Ct)_TEST4 (8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
134Activity_at_16uM_(delta-delta-Ct)_TEST4 (16μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
135Activity_at_35uM_(delta-delta-Ct)_TEST4 (35μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
136ACTIVITY_OUTCOME_TEST5The PubChem Activity Outcome for the subset of data included in the indicated Test number.Integer
137ACTIVITY_SCORE_TEST5The PubChem Activity Score for the subset of data included in the indicated Test number.Integer
138ASSAYDATA_COMMENT_TEST5Comments relevant to the subset of data included in the indicated Test number.String
139AbsAC40_Qualifier_TEST5<, =, or >String
140AbsAC40_uM_TEST5The concentration at which the fitted curve passes activity threshold nn.FloatμM
141pAbsAC40_M_TEST5Equal to -1*log10(AbsACnn).Float
142Hill_Slope_TEST5The slope at AC50Float
143S0_(delta-delta-Ct)_TEST5The fitted activity value at zero concentrationFloat%
144Sinf_(delta-delta-Ct)_TEST5The fitted activity value at infinite concentrationFloat%
145Num_Points_TEST5The number of data points used to generate the plotInteger
146Max_Activity_(delta-delta-Ct)_TEST5The maximum activity value observed, based on mean of replicates per concentrationFloat%
147Max_Activity_Conc_uM_TEST5The concentration at which the maximum activity is observedFloatμM
148Max_Concentration_uM_TEST5Maximum valid test concentrationFloatμM
149Activity_at_0.000060uM_(delta-delta-Ct)_TEST5 (6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
150Activity_at_0.00012uM_(delta-delta-Ct)_TEST5 (0.00012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
151Activity_at_0.00026uM_(delta-delta-Ct)_TEST5 (0.00026μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
152Activity_at_0.0005uM_(delta-delta-Ct)_TEST5 (0.0005μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
153Activity_at_0.001uM_(delta-delta-Ct)_TEST5 (0.001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
154Activity_at_0.0021uM_(delta-delta-Ct)_TEST5 (0.0021μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
155Activity_at_0.0042uM_(delta-delta-Ct)_TEST5 (0.0042μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
156Activity_at_0.008uM_(delta-delta-Ct)_TEST5 (0.008μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
157Activity_at_0.016uM_(delta-delta-Ct)_TEST5 (0.016μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
158Activity_at_0.03uM_(delta-delta-Ct)_TEST5 (0.03μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
159Activity_at_0.068uM_(delta-delta-Ct)_TEST5 (0.068μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
160Activity_at_0.135uM_(delta-delta-Ct)_TEST5 (0.135μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
161Activity_at_0.26uM_(delta-delta-Ct)_TEST5 (0.26μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
162Activity_at_0.5uM_(delta-delta-Ct)_TEST5 (0.5μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
163Activity_at_1uM_(delta-delta-Ct)_TEST5 (1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
164Activity_at_2.1uM_(delta-delta-Ct)_TEST5 (2.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
165Activity_at_4.2uM_(delta-delta-Ct)_TEST5 (4.2μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
166Activity_at_8uM_(delta-delta-Ct)_TEST5 (8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
167Activity_at_16uM_(delta-delta-Ct)_TEST5 (16μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
168Activity_at_35uM_(delta-delta-Ct)_TEST5 (35μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1 R03 DA031089-01

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