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Assay Detail

CHEMBL3706142

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Binding
Radioligand Competition Binding Assay: The affinity of the test compounds was determined by radioligand competition binding assay, using the known compound [3H]Ro-15-1788 (Flumazenil) (Perkin Elmer, 85.4 Ci/mmol) and the human recombinant GABA A receptor containing the alpha2, beta2, and gamma3 subunits.Membranes were prepared from HEK cells expressing hGABA A alpha2beta2- gamma3 receptor, and validated to ascertain protein concentration, receptor expression and to determine the Kd of the flumazenil as well as the Ki of a standard set of compounds before being used to test new compounds.The assay was carried out in 96 well plates; testing compounds using a 10 point semi-log dilution range from 19 uM top concentration. 100 ul of radioligand and 100 ul of membrane in 50 mM Tris-HCI and 0.05% F127 with 1 ul of test compound was incubated for 2 hours to allow the reaction to achieve equilibrium, and then harvested onto filter plates, dried and counted on a TopCount NXT. The data was analysed, and the Ki values were presented.
50
Total Activities
50
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Cell Type HEK293
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

Gamma-aminobutyric acid receptor subunit alpha-2 (CHEMBL4956)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Chemical compounds
(2015)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
Ki 50 7.41 8.29

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3647535 1 8.29
CHEMBL3647548 1 8.23
CHEMBL3647561 1 8.13
CHEMBL3647565 1 8.13
CHEMBL3647579 1 8.07
CHEMBL3647538 1 8.02
CHEMBL3647539 1 8.01
CHEMBL3647534 1 7.96
CHEMBL3647566 1 7.93
CHEMBL3647581 1 7.85
CHEMBL3647580 1 7.84
CHEMBL3647549 1 7.79
CHEMBL3647562 1 7.75
CHEMBL3647568 1 7.74
CHEMBL3647578 1 7.72
CHEMBL3647571 1 7.72
CHEMBL3647559 1 7.66
CHEMBL3647547 1 7.62
CHEMBL3647582 1 7.54
CHEMBL3647555 1 7.53
CHEMBL3647541 1 7.51
CHEMBL3647533 1 7.51
CHEMBL3647543 1 7.46
CHEMBL3647550 1 7.46
CHEMBL3647567 1 7.45
CHEMBL3647575 1 7.43
CHEMBL3647564 1 7.41
CHEMBL3647551 1 7.41
CHEMBL3647574 1 7.39
CHEMBL3647557 1 7.39

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3647535 Ki = 5.08 nM 8.29
CHEMBL3647548 Ki = 5.82 nM 8.23
CHEMBL3647565 Ki = 7.48 nM 8.13
CHEMBL3647561 Ki = 7.45 nM 8.13
CHEMBL3647579 Ki = 8.56 nM 8.07
CHEMBL3647538 Ki = 9.51 nM 8.02
CHEMBL3647539 Ki = 9.71 nM 8.01
CHEMBL3647534 Ki = 10.9 nM 7.96
CHEMBL3647566 Ki = 11.7 nM 7.93
CHEMBL3647581 Ki = 14.0 nM 7.85
CHEMBL3647580 Ki = 14.4 nM 7.84
CHEMBL3647549 Ki = 16.4 nM 7.79
CHEMBL3647562 Ki = 17.7 nM 7.75
CHEMBL3647568 Ki = 18.2 nM 7.74
CHEMBL3647571 Ki = 19.0 nM 7.72
CHEMBL3647578 Ki = 19.1 nM 7.72
CHEMBL3647559 Ki = 21.7 nM 7.66
CHEMBL3647547 Ki = 23.9 nM 7.62
CHEMBL3647582 Ki = 29.1 nM 7.54
CHEMBL3647555 Ki = 29.7 nM 7.53
CHEMBL3647541 Ki = 31.1 nM 7.51
CHEMBL3647533 Ki = 31.1 nM 7.51
CHEMBL3647543 Ki = 34.8 nM 7.46
CHEMBL3647550 Ki = 34.5 nM 7.46
CHEMBL3647567 Ki = 35.4 nM 7.45
CHEMBL3647575 Ki = 37.1 nM 7.43
CHEMBL3647551 Ki = 38.8 nM 7.41
CHEMBL3647564 Ki = 39.2 nM 7.41
CHEMBL3647574 Ki = 40.5 nM 7.39
CHEMBL3647557 Ki = 40.6 nM 7.39
CHEMBL3647572 Ki = 43.3 nM 7.36
CHEMBL3647569 Ki = 56.0 nM 7.25
CHEMBL3647563 Ki = 61.3 nM 7.21
CHEMBL3647573 Ki = 67.8 nM 7.17
CHEMBL3647560 Ki = 69.5 nM 7.16
CHEMBL3647544 Ki = 74.3 nM 7.13
CHEMBL3647577 Ki = 87.4 nM 7.06
CHEMBL3647558 Ki = 92.2 nM 7.04
CHEMBL3647552 Ki = 91.4 nM 7.04
CHEMBL3647545 Ki = 101.0 nM 7.00
CHEMBL3647570 Ki = 102.0 nM 6.99
CHEMBL3647537 Ki = 108.0 nM 6.97
CHEMBL3647553 Ki = 118.0 nM 6.93
CHEMBL3647546 Ki = 120.0 nM 6.92
CHEMBL3647542 Ki = 147.0 nM 6.83
CHEMBL3647576 Ki = 170.0 nM 6.77
CHEMBL3647540 Ki = 296.0 nM 6.53
CHEMBL3647554 Ki = 678.0 nM 6.17
CHEMBL3647556 Ki = 1750.0 nM 5.76
CHEMBL3647536 DARIGABAT Ki < 2.47 nM -