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BioAssay: AID 602311

Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in mouse brain membrane in vitro set 1

Name: Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in mouse brain membrane in vitro set 1. ..more
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 Tested Compounds
 Tested Compounds
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Active(26)
 
 
 Tested Substances
 Tested Substances
All(26)
 
 
Active(26)
 
 
AID: 602311
Data Source: The Scripps Research Institute Molecular Screening Center (DAGLB_INH_FLUO_1X%INH_SH_MBM_SET1)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network, Assay Provider
BioAssay Version:
Deposit Date: 2012-03-08
Hold-until Date: 2012-10-15
Modify Date: 2012-10-15

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 26
Related Experiments
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AIDNameTypeProbeComment
504411Fluorescence-based primary biochemical high throughput screening assay to identify inhibitors of human diacylglycerol lipase, beta (DAGLB)Screening depositor-specified cross reference: Primary screen (DAGLB inhibitors in singlicate)
504420Summary of the probe development efforts to identify inhibitors of human diacylglycerol lipase, beta (DAGLB)Summary3 depositor-specified cross reference: Summary (DAGLB inhibitors)
602403Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of recombinant antitarget DAGLa in vitroOther depositor-specified cross reference
602415Assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LC/MS-based biochemical inhibition of overexpressed DAGLb substrate turnover in vitroOther depositor-specified cross reference
624039Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of antitarget ABHD6 in vitro, set 2Confirmatory depositor-specified cross reference
624041Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse liver ABHD6 in vivoOther depositor-specified cross reference
624077Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse liver ABHD6 in vivo upon oral compound administrationOther depositor-specified cross reference
624468Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based biochemical dose response assayConfirmatory depositor-specified cross reference
624472Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of DAGLb by enantiomers of KT116Other depositor-specified cross reference
504445Fluorescence-based biochemical high throughput confirmation assay for inhibitors of human diacylglycerol lipase, beta (DAGLB)Screening same project related to Summary assay
602299Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of overexpressed DAGLb in vitro set 2Other same project related to Summary assay
602300Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of overexpressed DAGLb in vitro set 3Other same project related to Summary assay
602301Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of recombinant anti-target ABHD11 in vitro set 1Other same project related to Summary assay
602302Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of overexpressed DAGLb in vitro set 1Other same project related to Summary assay
602303Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of overexpressed DAGLb in vitro; triazole urea libraryOther same project related to Summary assay
602312Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of recombinant anti-target ABHD11 in vitro set 2Other same project related to Summary assay
602319Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of antitarget ABHD6 in vitroOther same project related to Summary assay
602320Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of DAGLb in vitroConfirmatory same project related to Summary assay
602321Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in mouse brain membrane in vitro set 3Other same project related to Summary assay
602322Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of antitarget ABHD6 in vitroConfirmatory same project related to Summary assay
602323Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in mouse brain membrane in vitro set 2Other same project related to Summary assay
602335Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of anti-target ABHD6 in situConfirmatory same project related to Summary assay
602337Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compoundsConfirmatory same project related to Summary assay
602339Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysisOther same project related to Summary assay
602341Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis for ABHD6Other same project related to Summary assay
602343Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse brain ABHD6 in vivo, set 2Other same project related to Summary assay
602345Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse macrophage DAGLb in vivoOther same project related to Summary assay
602347Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse brain ABHD6 in vivoOther same project related to Summary assay
602349Late stage assay provider results from the probe development effort to identify inhibitors of DAGLb: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) assay to distinguish systemic and peripheral inhibitorsOther same project related to Summary assay
602351Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based animal-based Activity-Based Protein Profiling (ABPP) MudPIT selectivity analysis for ABHD6Other same project related to Summary assay
602353Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based animal-based Activity-Based Protein Profiling (ABPP) MudPIT selectivity analysisOther same project related to Summary assay
602354Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical dose-response gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of DAGLb in situConfirmatory same project related to Summary assay
602355Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in vitroOther same project related to Summary assay
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Benjamin Cravatt, The Scripps Research Institute (TSRI)
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 DA025285
Grant Proposal PI: Benjamin Cravatt, The Scripps Research Institute (TSRI)
External Assay ID: DAGLB_INH_FLUO_1X%INH_SH_MBM_SET1

Name: Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of serine hydrolases in mouse brain membrane in vitro set 1.

Description:

Endocannabinoids (ECs) represent a unique group of lipids that function as chemical messengers in the nervous system. To date, the two principle ECs identified in mammals are N-arachidonoyl-ethanolamine (anandamide) and 2-arachidonoyl-glycerol (2-AG). They have been implicated in various physiological and pathological functions including appetite, pain, sensation, memory, and addiction (1). Unlike traditional neurotransmitters, which are stored in vesicles, ECs are synthesized and released on demand, and then rapidly degraded to terminate signaling. Thus, the metabolic pathways that govern EC turnover are critical in determining the magnitude and duration of neuronal signaling events (2). Endocannabinoid biosynthesis, in contrast to degradation, is poorly understood. Recently, two serine hydrolases, DAGL-a and -B, were cloned and found to selectively cleave sn-1 acyl chains from diacylglycerols (DAG) to generate 2-AG in vitro (3). Their function in the nervous system was validated in vivo by the generation of DAGL-a and -B knock-out mice (4, 5). However, it is still unclear to what extent DAGL-a/B catalytic activity contributes to 2-AG-mediated signaling. The development of potent and selective inhibitors would offer a means to perturb DAGL-a/B activity in a selective, reversible, and temporally-controlled manner. Given the non-selective nature of current DAGL-a/B inhibitors (6), specific chemical probes would serve as invaluable tools to delineate DAGL-a/B function in 2-AG signaling networks of the brain.

References:

1. Di Marzo, V. (2008) Targeting the endocannabinoid system: to enhance or reduce?, Nat Rev Drug Discov 7, 438-455.
2. Ahn, K., McKinney, M. K., and Cravatt, B. F. (2008) Enzymatic pathways that regulate endocannabinoid signaling in the nervous system, Chem Rev 108, 1687-1707.
3. Bisogno, T., Howell, F., Williams, G., Minassi, A., Cascio, M. G., Ligresti, A., Matias, I., Schiano-Moriello, A., Paul, P., Williams, E. J., Gangadharan, U., Hobbs, C., Di Marzo, V., and Doherty, P. (2003) Cloning of the first sn1-DAG lipases points to the spatial and temporal regulation of endocannabinoid signaling in the brain, J Cell Biol 163, 463-468.
4. Gao, Y., Vasilyev, D. V., Goncalves, M. B., Howell, F. V., Hobbs, C., Reisenberg, M., Shen, R., Zhang, M. Y., Strassle, B. W., Lu, P., Mark, L., Piesla, M. J., Deng, K., Kouranova, E. V., Ring, R. H., Whiteside, G. T., Bates, B., Walsh, F. S., Williams, G., Pangalos, M. N., Samad, T. A., and Doherty, P. (2010) Loss of Retrograde Endocannabinoid Signaling and Reduced Adult Neurogenesis in Diacylglycerol Lipase Knock-out Mice, J Neurosci 30, 2017-2024.
5. Tanimura, A., Yamazaki, M., Hashimotodani, Y., Uchigashima, M., Kawata, S., Abe, M., Kita, Y., Hashimoto, K., Shimizu, T., Watanabe, M., Sakimura, K., and Kano, M. (2010) The Endocannabinoid 2-Arachidonoylglycerol Produced by Diacylglycerol Lipase +/- Mediates Retrograde Suppression of Synaptic Transmission, Neuron 65, 320-327.
6. Hoover, H. S., Blankman, J. L., Niessen, S., and Cravatt, B. F. (2008) Selectivity of inhibitors of endocannabinoid biosynthesis evaluated by activity-based protein profiling, Bioorganic & Medicinal Chemistry Letters 18, 5838-5841.

Keywords:

late stage, late stage AID, assay provider, powders, counterscreen, diacylglycerol lipase, diacylglycerol lipase-beta, DAGL, DAGL-beta, DAGLB, hydrolase, serine hydrolase, appetite, pain, sensation, memory, addiction, activity-based protein profiling, ABPP, gel-based, activity-based probe, fluorophosphonate-rhodamine, FP-Rh, inhibitor, inhibition, serine hydrolases, mouse brain membrane, abhydrolase domain containing 6, ABHD6, acylpeptide hydrolase, APEH, fatty acid amide hydrolase, FAAH, arylacetamide deacetylase-like 1, KIAA1363, abhydrolase domain containing 12, ABHD12, monoglyceride lipase, MAGL, lysophospholipase 1, LYPLA1, lysophospholipase 2, LYPLA2, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Panel Information
Targets
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1ABHD626monoacylglycerol lipase ABHD6 [Mus musculus] [gi:31560264]
Taxonomy id: 10090
Gene id: 66082
2APEH125Acylpeptide hydrolase [Mus musculus] [gi:19343726]
Taxonomy id: 10090
Gene id: 235606
3FAAH1313fatty acid amide hydrolase [Mus musculus] [gi:123253900]
Taxonomy id: 10090
Gene id: 14073
4KIAA1363620neutral cholesterol ester hydrolase 1 [Mus musculus] [gi:30520239]
Taxonomy id: 10090
Gene id: 320024
5ABHD1226monoacylglycerol lipase ABHD12 [Mus musculus] [gi:159110817]
Taxonomy id: 10090
Gene id: 76192
6MAGL719monoacylglycerol lipase ABHD12 [Mus musculus] [gi:159110817]
Taxonomy id: 10090
Gene id: 23945
7LYPLA2224acyl-protein thioesterase 2 [Mus musculus] [gi:7242156]
Taxonomy id: 10090
Gene id: 26394
8LYPLA1224acyl-protein thioesterase 1 [Mus musculus] [gi:6678760]
Taxonomy id: 10090
Gene id: 18777

§ Panel component ID.
Protocol
Assay Overview:
The purpose of this assay is to assess test compound inhibition of serine hydrolases (SHs) in a complex proteome using a gel-based competitive activity-based proteomic profiling (ABPP) assay. In this assay, a complex proteome is incubated with test compound followed by reaction with a fluorescently-labeled serine hydrolase-specific activity-based probe, fluorophosphonate-rhodamine (FP-Rh). The reaction products are separated by SDS-PAGE and visualized in-gel using a flatbed fluorescence scanner. The percentage activity remaining is determined by measuring the integrated optical density (IOD) of the bands. As designed, test compounds that act as SH inhibitors will prevent enzyme-probe interactions, thereby decreasing the proportion of bound fluorescent probe, giving lower fluorescence intensity in the band in the gel.
Protocol Summary:
Mouse brain membrane proteome (25 uL of 1 mg/mL in Dulbecco's PBS [DPBS]) was treated with test compound (0.5 uL of a 50x stock in DMSO; 10 uM final concentration) or DMSO (0.5 uL) for 30 minutes at 37 C. The activity-based probe FP-Rh (0.5 uL of a 50x stock in DMSO; 1 uM final concentration) was added, and the reaction was incubated for 30 minutes at 37 C, quenched with an equal volume of 2x SDS-PAGE loading buffer (reducing), separated by SDS-PAGE, and visualized by in-gel fluorescent scanning. The percentage activity remaining for each SH was determined by measuring the integrated optical density of the individual protein bands relative to the DMSO-only (no compound) control. SHs with at least 50% inhibition by one or more compounds (abhydrolase domain containing 6 [ABHD6], acylpeptide hydrolase [APEH], fatty acid amide hydrolase [FAAH], arylacetamide deacetylase-like 1[KIAA1363], abhydrolase domain containing 12 [ABHD12], monoglyceride lipase [MAGL], lysophospholipase 1 [LYPLA1], and lysophospholipase 2 [LYPLA2]) are reported.
The percent inhibition for each compound was calculated as follows:
%_Inhibition = ( 1 - ( IOD_Test_Compound - Median_IOD_Low_Control ) / ( Median_IOD_High_Control - Median_IOD_Low_Control ) ) * 100
Where:
Test_Compound is defined as SH treated with test compound.
High_Control is defined as SH treated with DMSO only (no compound).
Low_Control is defined as background in a blank region of the gel.
PubChem Activity Outcome and Score:
The following applies to each panel in this assay:
Compounds with greater than or equal to 50% inhibition for a given SH were considered active. Compounds with less than 50% inhibition for a given SH were considered inactive.
The reported PubChem Activity Score has been normalized to 100% observed inhibition. Negative % inhibition values are reported as activity score zero.
ABHD6 Score: The PubChem Activity Score range for active compounds is 100-80. There are no inactive compounds.
APEH Score: The PubChem Activity Score range for active compounds is 100-100, and for inactive compounds 0-0.
FAAH Score: The PubChem Activity Score range for active compounds is 100-51, and for inactive compounds 45-0.
KIAA1363 Score: The PubChem Activity Score range for active compounds is 100-55, and for inactive compounds 40-0.
ABDH12 Score: The PubChem Activity Score range for inactive compounds is 100-0. There are no active compounds.
MAGL Score: The PubChem Activity Score range for active compounds is 100-63, and for inactive compounds 42-0.
LYPLA2 Score: The PubChem Activity Score range for active compounds is 100-50, and for inactive compounds 25-0.
LYPLA1 Score: The PubChem Activity Score range for active compounds is 100-50, and for inactive compounds 25-0.
Overall Outcome and Score:
Compounds active against at least one SH were considered active. Compounds inactive against all SHs were considered inactive.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Mouse brain membrane proteome (provided by Assay Provider)
FP-Rh (provided by Assay Provider)
DPBS (Cellgro 20-031-CV)
Comment
This assay was performed by the assay provider with powder samples of synthetic compounds.
Categorized Comment - additional comments and annotations
From BioAssay Depositor:
BAO: assay format: biochemical format: protein format: single protein format
BAO: bioassay specification: assay biosafety level: bsl1
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: bioassay specification: assay stage: secondary: selectivity
BAO: detection technology: fluorescence: fluorescence intensity
BAO: meta target detail: binding reporter specification: interaction: protein-small molecule
BAO: meta target: biological process target: regulation of molecular function
BAO: meta target: molecular target: protein target: enzyme: generic hydrolase
BAO: version: 1.4b1090
From PubChem:
Assay Format: Biochemical
Assay Test Type: In vitro
Result Definitions
Show more
TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Outcome [ABHD6]One of Active, Inactive, or Not Tested1monoacylglycerol lipase ABHD6 [Mus musculus]Outcome
2Score [ABHD6]BioAssay Score.1Integer
3Inhibition at 10 uM [ABHD6] (10μM**)Inhibition of endogenous mouse ABHD6 upon 10 uM compound treatment as assessed by gel-based competitive ABPP.1Integer%
4Outcome [APEH]One of Active, Inactive, or Not Tested2Acylpeptide hydrolase [Mus musculus]Outcome
5Score [APEH]BioAssay Score.2Integer
6Inhibition at 10 uM [APEH] (10μM**)Inhibition of endogenous mouse APEH upon 10 uM compound treatment as assessed by gel-based competitive ABPP.2Integer%
7Outcome [FAAH]One of Active, Inactive, or Not Tested3fatty acid amide hydrolase [Mus musculus]Outcome
8Score [FAAH]BioAssay Score.3Integer
9Inhibition at 10 uM [FAAH] (10μM**)Inhibition of endogenous mouse FAAH upon 10 uM compound treatment as assessed by gel-based competitive ABPP.3Integer%
10Outcome [KIAA1363]One of Active, Inactive, or Not Tested4neutral cholesterol ester hydrolase 1 [Mus musculus]Outcome
11Score [KIAA1363]BioAssay Score.4Integer
12Inhibition at 10 uM [KIAA1363] (10μM**)Inhibition of endogenous mouse KIAA1363 upon 10 uM compound treatment as assessed by gel-based competitive ABPP.4Integer%
13Outcome [ABDH12]One of Active, Inactive, or Not Tested5monoacylglycerol lipase ABHD12 [Mus musculus]Outcome
14Score [ABDH12]BioAssay Score.5Integer
15Inhibition at 10 uM [ABDH12] (10μM**)Inhibition of endogenous mouse ABHD12 upon 10 uM compound treatment as assessed by gel-based competitive ABPP.5Integer%
16Outcome [MAGL]One of Active, Inactive, or Not Tested6monoacylglycerol lipase ABHD12 [Mus musculus]Outcome
17Score [MAGL]BioAssay Score.6Integer
18Inhibition at 10 uM [MAGL] (10μM**)Inhibition of endogenous mouse MAGL upon 10 uM compound treatment as assessed by gel-based competitive ABPP.6Integer%
19Outcome [LYPLA2]One of Active, Inactive, or Not Tested7acyl-protein thioesterase 2 [Mus musculus]Outcome
20Score [LYPLA2]BioAssay Score.7Integer
21Inhibition at 10 uM [LYPLA2] (10μM**)Inhibition of endogenous mouse LYPLA2 upon 10 uM compound treatment as assessed by gel-based competitive ABPP.7Integer%
22Outcome [LYPLA1]One of Active, Inactive, or Not Tested8acyl-protein thioesterase 1 [Mus musculus]Outcome
23Score [LYPLA1]BioAssay Score.8Integer
24Inhibition at 10 uM [LYPLA1] (10μM**)Inhibition of endogenous mouse LYPLA1 upon 10 uM compound treatment as assessed by gel-based competitive ABPP.8Integer%

** Test Concentration. § Panel component ID.
Additional Information
Grant Number: 1 R01 DA025285

Classification
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