Biology Department of WuXi AppTec (Shanghai) Co., Ltd.
ADORA2A (Adenosine A2A Receptor)
Cell Line Name: HEK293/A2A/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADORA2A ; A2aR; ADORA2; RDC8 Accession Number: NM_000675 G coupling Gs
Functional Assay-Calcium Flux Assay
ADORA2A NECA 120 Agonist NECA CPA 100 Measurement: Intracellular Ca2+ 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40
Activity (%) Activity 20 Agonist effect EC50: 5.4 nM 0 -20 -2 0 2 4 6 Log [Compound] nM ADORA2A 120 Antagonist CGS-15943 CGS-15943 100 2+ ZM241385 Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 20
Inhibition (%) Inhibition Stimulant NECA: 40 nM 0 Antagonist effect IC50: 0.22 nM -20 -4 -2 0 2 4 Log [Compound] nM
ADORA2A (Adenosine A2A Receptor)
Cellular Background: HEK293 Species/Source: Human/Recombinant Gene Synonyms: ADORA2A ; A2aR; ADORA2; RDC8 Accession Number: NM_000675
Radio ligand binding Assay
ADORA2A Ligand [3H] CGS 21680 125 NECA Ligand concentration 6 nM 100 CPA ZM241385 Kd (nM) 47.5 nM 75 CGS15943 Non specific NECA (10 μM) 50
Incubation 120 min/RT Inhibition % 25
Detection method Scintillation counting 0
Reference: NECA (IC50:82.0 nM) -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM
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Alpha1A (Alpha-1-adrenergic receptor)
Cell Line Name: HEK293/Alpha1A Species/Source: Human/Recombinant Gene Synonyms: ADRA1A; ADRA1C; ADRA1L1; ALPHA1AAR Accession Number: NM_000680 (isoform 1) G coupling Gq
Functional Assay-Calcium Flux Assay
Alpha1A 120 Agonist (-)Epinephrine (-)-Epinephrine (E4642) 100 A-61603 2+ Measurement: Intracellular Ca 80 60 Detection method: FLIPR Fluorimetry 40
(%) Activity Incubation 25℃ 20 0 Agonist effect EC50: 4.0 nM -2 -1 0 1 2 3 4 Log [Compound] nM
Alpha1A 5-Methylurapidil Antagonist 5-Methylurapidil 120 WB-4104 Measurement: Intracellular Ca2+ 100 Prazosin 80 BMY7378 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 Inhibition (%) Inhibition 20 Stimulant (-)Epinephrine : 6.3 nM 0 -2 0 2 4 6 Antagonist effect IC50: 3.6 nM Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]prazosin Alpha1A 125 WB4101 Ligand concentration 0.2 nM 100 Epinephrine Prazosin Kd (nM) 0.09 nM 75 Non specific epinephrine (0.1 mM) 50
% Inhibition % 25 Incubation 60 min/RT 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: WB 4101 (IC50:0.28 Log [Compound] nM nM)
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Alpha2A (Alpha-2-adrenergic receptor)
Cell Line Name: HEK293/Alpha2A/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRA2A; ADRA2; ADRAR; ZNF32; ADRA2R; ALPHA2AAR Accession Number: NM_000681 (isoform 1) G coupling Gi
Functional Assay-Calcium Flux Assay
Alpha2A UK 14304 Agonist UK14304 120 Epinephrine (E4250) 100 Measurement: Intracellular Ca2+ 80 Detection method: FLIPR Fluorimetry 60 ℃ 40 Incubation 25 (%) Activity 20
Agonist effect EC50: 0.14 nM 0 -20 -2 0 2 Log [Compound] nM
Alpha2A 125 Antagonist Yohimbine hydrochloride Yohimbin 100 RX 821002 2+ Measurement: Intracellular Ca 75 Prazosin Detection method: FLIPR Fluorimetry 50 25
Incubation 25℃ (%) Inhibition 0
Stimulant UK14304 : 4.3 nM -25 -2 -1 0 1 2 3 4 Antagonist effect IC50: 32.6 nM Log [Compound] nM
Radio ligand binding Assay
Alpha2A Ligand [3H]RX 821002 125 Epinephrine Ligand concentration 0.5 nM 100 Prazosin Yohimbin 75 Kd (nM) 0.46 nM RX821002 Non specific epinephrine (0.1 mM) 50 UK14304
Incubation 60 min/RT Inhibition % 25 0 Detection method Scintillation counting -2 -1 0 1 2 3 4 5 Reference: Rx81002(IC50:2.3 nM) Log [Compound] nM
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Beta1 (Beta-1 Adrenergic Receptor)
Cell Line Name: HEK293/Beta1/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRB1; RHR; B1AR; ADRB1R; BETA1AR Accession Number: NM_000684 G coupling Gs
Functional Assay-Calcium Flux Assay
Beta1 (-)Epinephrine 120 Isoproterenol Agonist (-)Epinephrine 100 Salbutamol hemisulfate Measurement: Intracellular Ca2+ 80 60 Detection method: FLIPR Fluorimetry 40
Activity (%) Activity Incubation 25℃ 20 0 Agonist effect EC50: 2.72 nM -20 -2 -1 0 1 2 3 4 5 Log [Compound] nM
Beta1 120 Antagonist Carvedilol Carvedilol 100 (± )-Propranolol hydrochloride Measurement: Intracellular Ca2+ 80 Atenolol 60 ICI 118 551 hydrochloride Detection method: FLIPR Fluorimetry 40 20
Inhibition (%) Inhibition Incubation 25℃ 0 -20 Stimulant (-)-Epinephrine : 20 nM -2 -1 0 1 2 3 4 Log [Compound] nM Antagonist effect IC50: 4.75 nM
Radio ligand binding Assay
Beta1 125 Ligand [3H](-)CGP 12177 Atenolol
Ligand concentration 0.3 nM 100 Salbutamol Propranolol 75 Kd (nM) 0.04 nM ICI118551 50 Non specific propranolol (2 µM) Isoproternol
% Inhibition % 25 Incubation 120 min/RT 0 Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Reference: Atenolol (IC50:857 nM) Log [Compound] nM
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Beta2 (Beta-2 Adrenergic Receptor)
Cell Line Name: HEK293/Beta2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRB2; BAR; B2AR; ADRBR; ADRB2R; BETA2AR Accession Number: NM_000024 G coupling Gs/Gi
Functional Assay-Calcium Flux Assay
Beta2 Adrenaline Agonist Adrenaline 120 Isoproterenol 100 Salbutamol hemisulfate 2+ Measurement: Intracellular Ca 80
Detection method: FLIPR Fluorimetry 60 40
Incubation 25℃ (%) Activity 20
Agonist effect EC50: 0.11 nM 0 -20 -4 -3 -2 -1 0 1 2 3 4 Log [Compound] nM
Beta2 Carvedilol Antagonist Carvedilol 125 (± )-Propranolol hydrochloride ICI 118551 Measurement: Intracellular Ca2+ 100 75 Detection method: FLIPR Fluorimetry 50
Incubation 25℃ 25
Inhibition (%) Inhibition Stimulant Adrenaline: 0.43 nM 0 -25 Antagonist effect IC50: 7.07 nM -2 -1 0 1 2 3 Log [Compound] nM
Radio ligand binding Assay
Beta2 Ligand [3H](-)CGP 12177 125 Atenolol Ligand concentration 0.3 nM 100 Salbutamol Propranolol Kd (nM) 0.1 nM 75 ICI118551 Non specific propranolol (2 µM) 50 Isoproternol
Incubation 120 min/RT Inhibition % 25
Detection method Scintillation counting 0
Reference: ICI118551(IC50:3.3 nM) -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM
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CB1 (Cannabinoid Type 1 Receptor)
Cell Line Name: CHO/CB1/Gα15 Species/Source: Human/Recombinant Gene Synonyms: CNR1; CB1; CNR; CB-R; CB1A; CB1R; CANN6; CB1K5 Accession Number: NM_016083 (isoform1) G coupling Gi
Functional Assay-Calcium Flux Assay
CB1 WIN55212-2 Agonist WIN55212-2 120 CP55940 2+ 100 Anandamide Measurement: Intracellular Ca 80
Detection method: FLIPR Fluorimetry 60 40
Incubation 25℃ (%) Activity 20
Agonist effect EC50:352 nM 0 -20 -1 0 1 2 3 4 5 6 Log [Compound] nM
CB1 Antagonist Rimonabant hydrochloride Rimonabant 120 2+ AM281 Measurement: Intracellular Ca 100 AM630 Detection method: FLIPR Fluorimetry 80 60 Incubation 25℃ 40 20 Inhibition (%) Inhibition Stimulant WIN55212-2: 4000 nM 0 -20 Antagonist effect IC50: 12.5 nM -1 0 1 2 3 4 5 6 log [Con.] nM
CB1 (Cannabinoid Type 1 Receptor)
Cellular Background: CHO-K1 Species/Source: Human/Recombinant Gene Synonyms: CNR1; CB1; CNR; CB-R; CB1A; CB1R; CANN6; CB1K5 Accession Number: X54937
Radio ligand binding Assay
Ligand [3H]CP 55940 CB1 125 Ligand concentration 0.4 nM CP55940 100 WIN55212-2 AM281 Kd (nM) 3.7 nM 75 Anandamide Non specific WIN 55212-2 (5 µM) 50
Incubation 120 min/37℃ Inhibition % 25 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: CP 55940 (0.81 nM) Log [Compound] nM
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CB2 (Cannabinoid Type 2 Receptor)
Cell Line Name: CHO/CB2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: CNR2; CB2; CX5; CB-2 Accession Number: NM_001841 G coupling Gi
Functional Assay-Calcium Flux Assay
CB2 120 Agonist CP55940 CP55940 2+ 100 WIN55215-2 Measurement: Intracellular Ca 80 Anandamide Detection method: FLIPR Fluorimetry 60 40
℃ Activity (%) Incubation 25 20 Agonist effect EC50: 199 nM 0 -1 0 1 2 3 4 5 Log [Compound] nM
Antagonist AM630 CB2 120 2+ AM630 Measurement: Intracellular Ca 100 AM281 Detection method: FLIPR Fluorimetry 80 Rimonbant 60 Incubation 25℃ 40 20 Stimulant CP55940: 300 nM (%) Inhibition 0 Antagonist effect IC50: 563 nM -20 0 1 2 3 4 5 Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]CP 55940 CB2 125 Ligand concentration 0.5 nM CP55940 100 WIN55212-2 Anandomide Kd (nM) 5.1 nM 75 AM630 Non specific WIN 55212-2 (5 µM) 50 AM281
% Inhibition % 25 Incubation 120 min/37℃ 0
Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: WIN55212-2(IC50: Log [Compound] nM 2.2nM)
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CCKA (Cholecystokinin A Receptor)
Cell Line Name: HEK293/CCKA Species/Source: Human/Recombinant Gene Synonyms: CCKAR; CCK-A; CCK1R; CCKRA; CCK1-R Accession Number: NM_000730 G coupling Gq
Functional Assay-Calcium Flux Assay
CCKA Agonist CCK-4 120 CCK-4 2+ Measurement: Intracellular Ca 100 CCK-8s 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 (%) Activity Agonist effect EC50: 80 nM 20 0 -4 -2 0 2 4 Log [Compound] nM CCKA Antagonist Lorglumide Lorglumide 120 2+ Devazepide Measurement: Intracellular Ca 100 LY 225910 Detection method: FLIPR Fluorimetry 80 60 Incubation 25℃ 40 20 Stimulant CCK-4: 557 nM (%) Inhibition 0 Antagonist effect IC50: 635 nM -20 0 1 2 3 4 5 Log [Compound] nM CCKA (Cholecystokinin A Receptor)
Cellular Background: 1321-N1 Species/Source: Human/Recombinant Gene Synonyms: CCKAR; CCK-A; CCK1R; CCKRA; CCK1-R Accession Number: L19315
Radio ligand binding Assay
Ligand [3H]CCK-8s CCKA 125 CCK8s Ligand concentration 0.5 nM 100 Devazepide LY225910 Kd (nM) 0.22 nM 75 50 Non specific CCK-8s (1 μM)
% Inhibition % 25
Incubation 60 min/RT 0
Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM Reference: CCK-8s(IC50: 2.4 nM)
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D1 (Dopamine D1 Receptor)
Cell Line Name: HEK293/D1/Gα15 Species/Source: Human/Recombinant Gene Synonyms: DRD1; DADR; DRD1A Accession Number: NM_000794 G coupling Gs
Functional Assay-Calcium Flux Assay
D1 (± )-SKF-38393 120 Agonist (±)-SKF-38393 Dopamine 100 2+ 7-OH-DPAT Measurement: Intracellular Ca 80
Detection method: FLIPR Fluorimetry 60 40
Incubation 25℃ (%) Activity 20
Agonist effect EC50: 5.1 nM 0 -20 -2 0 2 4 6 Log [Compound] nM
D1 Antagonist R(+)-SCH-23390 125 L-745870 hydrochloride 100 SCH-23390 75 2+ Measurement: Intracellular Ca 50 Detection method: FLIPR Fluorimetry 25 0
Inhibition (%) Inhibition Incubation 25℃ -25 -2 0 2 4 6 -50 Stimulant (±)-SKF-38393: 11 nM Log [Compound] nM Antagonist effect IC50: 1.3 nM
Radio ligand binding Assay
D1 125 Ligand [3H]SCH 23390 Dopamine 100 7-OH-DPAT Ligand concentration 0.3 nM L-745870 75 Kd (nM) 0.6 nM Droperidol 50 R(+)-SCH23390 Non specific droperidol (10 µM)
% Inhibition % 25
Incubation 60 min/RT 0
Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM Reference: R(+)-SCH23390(IC50:1.7 nM)
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D2 (Dopamine D2 Receptor)
Cell Line Name: HEK293/D2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: DRD2; D2R; D2DR Accession Number: NM_000795 G coupling Gi
Functional Assay-Calcium Flux Assay
D2 Dopamine 120 7-OH-DPAT Agonist Dopamine 100
Measurement: Intracellular Ca2+ 80 60 Detection method: FLIPR Fluorimetry 40
(%) Activity Incubation 25℃ 20 0 Agonist effect EC50: 3.1 nM -20 -2 -1 0 1 2 3 4 Log [Compound] nM
D2 120 Amisulpride Antagonist Amisulpride Droperidol 100 Measurement: Intracellular Ca2+ (+)Butaclamol 80 (-)Butaclamol Detection method: FLIPR Fluorimetry 60 40 Incubation 25℃ Inhibition% 20
Stimulant Dopamine: 10.5 nM 0 -2 0 2 4 6 Antagonist effect IC50: 4.8 nM Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]7-OH-DPAT D2 125 Dopamine Ligand concentration 1 nM 100 7-OH-DPAT L-745870 Kd (nM) 1.7 nM 75 Droperidol Non specific droperidol (10 µM) 50 R(+)-SCH23390 (+)-Butaclamol Incubation 60 min/RT Inhibition % 25 0 Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Reference: (+)-Butaclamol(IC50:1.4 Log [Compound] nM nM)
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Eta (Endothelin Receptor Type A)
Cell Line Name: HEK293/ETA Species/Source: Human/Recombinant Gene Synonyms: EDNRA; ETA; ET-A; ETAR; ETRA; MFDA; ETA-R; hET-AR Accession Number: NM_001957 G coupling Gq
Functional Assay-Calcium Flux Assay
Eta 125 Agonist Endothelin 1 Endothelin 1 Measurement: Intracellular Ca2+ 100 Endothelin 3 75 Detection method: FLIPR Fluorimetry 50 ℃
Incubation 25 % Activity 25 Agonist effect EC50: 1.6 nM 0 -2 -1 0 1 2 3 Log [Compound] nM
Eta BQ-123 Antagonist BQ-123 125 BQ-788 100 Measurement: Intracellular Ca2+ 75 Detection method: FLIPR Fluorimetry 50
Incubation ℃ 25 25 (%) Inhibition 0 Stimulant Endothelin 1: 3.2 nM -25 Antagonist effect IC50: 3.0 nM -3 -2 -1 0 1 2 3 4 Log [Compound] nM
Radio ligand binding Assay
ETa Ligand [125I]endothelin-1 125 Ligand concentration 0.03 nM Endothelin-1 100 BQ-123 BQ-788 Kd (nM) 0.03 nM 75 Non specific endothelin-1 (0.1 μM) 50
Incubation 120 min/37°C Inhibition % 25 Detection method Scintillation counting 0 -5 -4 -3 -2 -1 0 1 2 3 4 Reference: endothelin-1(IC50:0.12 Log [Compound, nM] nM)
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H1 (Histamine H1 receptor)
Cell Line Name: HEK293/H1 Species/Source: Human/Recombinant Gene Synonyms: HRH1; H1R; H1-R; HH1R; hisH1 Accession Number: NM_000861 G coupling Gq
Functional Assay-Calcium Flux Assay
H1 histamine 125 2-PEA Agonist Histamine 100 2+ Measurement: Intracellular Ca 75
Detection method: FLIPR Fluorimetry 50
Incubation 25℃ % Activity 25 0 Agonist effect EC50: 2.39 nM -25 -2 0 2 4 Log [Compound] nM Antagonist Pyrilamine maleate salt H1 125 Measurement: Intracellular Ca2+ Pyrilamine 100 Clobenpropit Detection method: FLIPR Fluorimetry 75 Cimetidine Incubation 25℃ 50 25
Stimulant Histamine: 6.3 nM (%) Inhibition 0 Antagonist effect IC50: 3.87 Nm -25 -2 0 2 4 6 Log [Compound] nM
Radio ligand binding Assay
H1 Ligand [3H]pyrilamine 125 Pyrilamine Ligand concentration 1 nM 100 Histamine Clobenpropit 75 Kd (nM) 2.3 nM Cimetdine Non specific Pyrilamine (1 µM) 50
% Inhibition % 25 Incubation 60 min/RT 0
Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Reference: Pyrilamine(IC50:6.8nM) Log [Compound] nM
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Cell Line Name: HEK293/H2 Species/Source: Human/Recombinant Gene Synonyms: H2; HRH2; H2R Accession Number: NM_022304 G coupling Gq
Functional Assay-Calcium Flux Assay
H2 Imetit Agonist Amthamine 120 Amthamine 100 dihydrobromide Histamine 80 2+ Measurement: Intracellular Ca 60 40 Detection method: FLIPR Fluorimetry (%) Activity 20 Incubation 25℃ 0 -20 Agonist effect EC50: 1125 nM 0 1 2 3 4 5 Log [Compound] nM
H2 Cimetidine Antagonist Cimetidine 125 Ratinidine Measurement: Intracellular Ca2+ 100 75 Detection method: FLIPR Fluorimetry 50 Incubation 25℃
%inhibition 25 Stimulant Amthamine 0 dihydrobromide: 8000 nM 0 1 2 3 4 5 Antagonist effect IC50: 111.8 nM Log [Compound] nM
Radio ligand binding Assay
H2 Ligand [3H]Tiotidine 125 Histamine Ligand concentration 1 nM 100 Clobenpropit Cimetdine Kd (nM) 0.8 nM 75 Imetit Non specific Cimetdine(10uM) 50
Incubation 60 min/RT Inhibition % 25
Detection method Scintillation counting 0
Reference: Cimetdine(IC50:472.7nM) -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM
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5HT1A (5-HT1A Receptor)
Cell Line Name: HEK293/5-HT1A/Gα15 Species/Source: Human/Recombinant Gene Synonyms: HTR1A; G-21; 5HT1a; PFMCD; 5-HT1A; 5-HT-1A; ADRBRL1; ADRB2RL1 Accession Number: NM_000524 G coupling Gi
Functional Assay-Calcium Flux Assay
Agonist 5-CT 5HT1A 120 5-CT 2+ Serotonin Measurement: Intracellular Ca 100 8-OH-DPAT Detection method: FLIPR Fluorimetry 80 60 Incubation 25℃ 40
Agonist effect EC50: 0.72 nM (%) Activity 20 0 -20 -3 -2 -1 0 1 2 3 4 5 Antagonist Methiothepin mesylate salt Log [Compound] nM 5HT1A Methiothepin 2+ Measurement: Intracellular Ca 125 WAY-100635 Ketanserin Detection method: FLIPR Fluorimetry 100 Incubation 25℃ 75 Stimulant 5-CT: 20 nM 50 25
Inhibition (%) Inhibition Antagonist effect IC50: 42.3 nM 0 0 2 4 6 -25 Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]8-OH-DPAT 5HT1A 125 Ligand concentration 0.3 nM 5-CT 100 Serotonin 8-OH-DPAT Kd (nM) 0.6 nM 75 WAY-100635 Non specific 8-OH-DPAT (10 µM) 50 Ketanserin
Incubation 60 min/RT Inhibition % 25 0
Detection method Scintillation counting -5 -4 -3 -2 -1 0 1 2 3 4 Reference: 8-OH-DPAT(IC50:1.6nM) Log [Compound] nM
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5HT1B (5-HT1B Receptor)
Cell Line Name: CHO-K1/5-HT1B Species/Source: Human/Recombinant Gene Synonyms: 5-HT-1B; 5-HT-1D-beta; 5-HT1B; 5-HT1DB; HTR1D2; HTR1DB; S12 Accession Number: NM_000863 G coupling Gi
Functional Assay-Calcium Flux Assay
Agonist 5-HT 5-HT1B 120 5-HT Measurement: cAMP Assay 100 5-CT 80 Detection method: HTRF 8-OH-DPAT 60 Incubation 25℃ 40 20 Agonist effect EC50: 3.75 nM Activity % 0 -20 -40 -2 0 2 4 6 Log [Compound] nM
5-HT1B Antagonist GR127935 140 GR127935 Measurement: cAMP Assay 120 Yohimbine hydrochloride 100 Detection method: HTRF Ketanserin 80 Incubation 25℃ 60 40
Stimulant 5-HT: 20 nM Inhibition % 20 Antagonist effect IC50: 0.92 nM 0 -20 -2 0 2 4 6 Log [Compound] nM
5HT1B (5-HT1B Receptor)
Cellular Background: Chem-1 Species/Source: Human/Recombinant Gene Synonyms: 5-HT-1B; 5-HT-1D-beta; 5-HT1B; 5-HT1DB; HTR1D2; HTR1DB; S12 Accession Number: NM_000863
Radio ligand binding Assay
Ligand [3H]GR125743 5HT1B 125 Ligand concentration 4 nM Serotonin 100 Ketanserin Kd (nM) 4.7 nM 8-OH-DPAT 75 Non specific Serotonin (10 μM) GR127935 50 Incubation 120 min/RT
% Inhibition % 25
Detection method Scintillation counting 0
Reference: Serotonin(IC50:85.4 nM) -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM
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5HT2A (5-HT2A Receptor)
Cell Line Name: HEK293/5-HT2A Species/Source: Human/Recombinant Gene Synonyms: HTR2A; HTR2; 5-HT2A Accession Number: NM_000621 G coupling Gq
Functional Assay-Calcium Flux Assay
Agonist 5-CT 5HT2A Measurement: Intracellular Ca2+ 120 5-CT 100 Serotonin (± )-DOI hydrochloride 80 Detection method: FLIPR Fluorimetry 5-Methoxytryptamine 60 Incubation 25℃ 40
Agonist effect EC50: 25.5 nM (%) Activity 20 0 -20 -4 -2 0 2 4 Log [Compound] nM Antagonist Methiothepin mesylate salt 5HT2A 125 2+ Methiothepin mesylate salt Measurement: Intracellular Ca 100 Ketanserin Detection method: FLIPR Fluorimetry 75 50
Incubation 25℃ 25
Inhibition (%) Inhibition Stimulant 5-CT: 47 nM 0 -25 Antagonist effect IC50: 138.5 nM -2 0 2 4 6 Log [Compound] nM
5HT2A (5-HT2A Receptor)
Cellular Background: CHO-K1 Species/Source: Human/Recombinant Gene Synonyms: HTR2A; HTR2; 5-HT2A Accession Number: X57830
Radio ligand binding Assay
Ligand [3H]-Ketanserin 5HT2A 125 Ligand concentration 0.5 nM (±)DOI 100 5-Methoxytryptamine Serotonin Kd (nM) 0.6 nM 75 BW723C86 Non specific (±)DOI (1 µM) 50 Ketanserin SB206553 Incubation 60 min/RT Inhibition % 25 0 Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Reference: (±)DOI (IC50:41.0nM) Log [Compound] nM
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5HT2B (5-HT2B Receptor)
Cell Line Name: HEK293/5-HT2B Species/Source: Human/Recombinant Gene Synonyms: 5-HT (2B); 5-HT-2B; 5-HT2B Accession Number: NM_000867 G coupling Gq
Functional Assay-Calcium Flux Assay
5HT2B 125 Agonist BW-723C86 Methiothepin 100 Measurement: Intracellular Ca2+ 75 Detection method: FLIPR Fluorimetry 50 Incubation 25℃
% Inhibition % 25
Agonist effect EC50: 4.827 nM 0
-2 -1 0 1 2 3 4 5 Log [Compound] nM 5HT2B 125 Antagonist Methiothepin mesylate salt BW-723C86 100 Measurement: Intracellular Ca2+ 75
Detection method: FLIPR Fluorimetry 50
Incubation 25℃ Activity % 25 Stimulant BW-723C86: 20 nM 0
Antagonist effect IC50: 34.6 nM -2 -1 0 1 2 3 4 5 Log [Compound] nM
5HT2B (5-HT2B Receptor)
Cellular Background: CHO-K1 Species/Source: Human/Recombinant Gene Synonyms: 5-HT (2B); 5-HT-2B; 5-HT2B Accession Number: X77307
Radio ligand binding Assay
Ligand [3H]LSD 5HT2B 125 Ligand concentration 1 nM (±)DOI 100 5-Methoxytryptamine Kd (nM) 0.93 nM Serotonin 75 Non specific Serotonin (50 μM) BW723C86 50 Ketanserin Incubation 60 min/RT SB206553
% Inhibition % 25
Detection method Scintillation counting 0
Reference: (±)DOI (IC50: 34.1 nM) -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM
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M1 (Muscarinic Acetylcholine M1 Receptor)
Cell Line Name: HEK293/M1 Species/Source: Human/Recombinant Gene Synonyms: CHRM1; M1; HM1; M1R Accession Number: NM_000738 G coupling Gq/Gi/Gs
Functional Assay-Calcium Flux Assay
M1 Arecaidine propargylester 120 Agonist Arecaidine propargyl Acetylcholine 100 Carbachol ester 80 60 Measurement: Intracellular Ca2+ 40 Detection method: FLIPR Fluorimetry (%) Activity 20 0 Incubation 25℃ -20 -3 -2 -1 0 1 2 3 4 Agonist effect EC50: 0.95 nM Log [Compound] nM
M1 Atropine Antagonist Pirenzepine 125 Pirenzepine dihydrochloride 100 Methoctramine 4-DAMP Measurement: Intracellular Ca2+ 75 50
Detection method: FLIPR Fluorimetry 25
Inhibition% Incubation 25℃ 0 -2 0 2 4 6 Stimulant Arecaidine -25 log [Con.] nM propargylester : 1.4 nM Antagonist effect IC50: 34 nM
Radio ligand binding Assay
M1 Ligand [3H]NMS 125 Atropine Ligand concentration 0.2 nM 100 4-DAMP Pirenzepine Kd (nM) 0.07 nM 75 Methocatramine Non specific Atropine (1 μM) 50
Incubation 60 min/RT Inhibition% 25
Detection method Scintillation counting 0
Reference: Pirenzepine (IC50:13.2 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM nM)
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M2 (Muscarinic Acetylcholine M2 Receptor)
Cell Line Name: HEK293/M2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: CHRM2; M2; HM2 Accession Number: NM_000739 G coupling Gi
Functional Assay-Calcium Flux Assay
M2
Agonist Arecaidine propargyl 120 APL 100 Carbachol ester 80 Acetylcholine
Measurement: Intracellular Ca2+ 60 40
Detection method: FLIPR Fluorimetry (%) Activity 20
Incubation 25℃ 0 -20 Agonist effect EC50: 25.2 nM -2 -1 0 1 2 3 4 5 Log [Compound] nM
M2 Atropine 125 Pirenzepoine Antagonist Atropine 100 Methoctramine Measurement: Intracellular Ca2+ 75 4-DAMP 50 Detection method: FLIPR Fluorimetry 25
Activity (%) Activity Incubation 25℃ 0 Stimulant Arecaidine -25 -2 0 2 4 6 propargylester : 89 nM Log [Compound] nM Antagonist effect IC50: 0.14 nM
Radio ligand binding Assay
M2 Ligand [3H]NMS 125 Atropine Ligand concentration 0.2 nM 100 4-DAMP Pirenzepine Kd (nM) 0.19 nM 75 Methoctramine Non specific Atropine (1 μM) 50
Inhibition% Incubation 60 min/RT 25 0 Detection method Scintillation counting
Reference: Methoctramine (IC50:1.4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM nM)
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M3 (Cholinergic/Acetylcholine M3 Receptor)
Cell Line Name: HEK293/M3 Species/Source: Human/Recombinant Gene Synonyms: CHRM3; M3; HM3; EGBRS Accession Number: NM_000740 G coupling Gq
Functional Assay-Calcium Flux Assay
M3
120 Arecaidine propargylester Agonist Arecaidine propargyl 100 Carbachol
ester 80 Acetylcholine 60 Measurement: Intracellular Ca2+ 40 Detection method: FLIPR Fluorimetry (%) Activity 20 0 Incubation ℃ 25 -20 -4 -2 0 2 4 Agonist effect EC50: 0.36 nM Log [Compound] nM
M3 Atropine Antagonist Atropine 125 4-DAMP Pirenzepine Measurement: Intracellular Ca2+ 100 Detection method: FLIPR Fluorimetry 75 50 Incubation 25℃ 25
Inhibition (%) Inhibition Stimulant Arecaidine 0 propargylester : 1.0 nM -25 -4 -2 0 2 4 6 Antagonist effect IC50: 1.4 nM Log [Compound] nM
Radio ligand binding Assay
M3 125 Ligand [3H]NMS Atropine 100 Ligand concentration 0.2 nM 4-DAMP Pirenzepine 75 Kd (nM) 0.18 nM Methocatramine 50 Non specific Atropine (1 μM) Inhibition% 25 Incubation 60 min/RT 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: 4-DAMP (IC50:1.1 nM) Log [Compound] nM
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op-delta (δ-opioid Receptor)
Cell Line Name: HEK293/delta/Gα15 Species/Source: Human/Recombinant Gene Synonyms: OPRD1; OPRD Accession Number: NM_000911 G coupling Gi
Functional Assay-Calcium Flux Assay
op-delta DPDPE 120 DAMGO Agonist DPDPE 100 U-50488 Measurement: Intracellular Ca2+ 80 60 Detection method: FLIPR Fluorimetry 40 Incubation 25℃ (%) Activity 20 0 Agonist effect EC50: 0.5 nM -20 -2 0 2 4 6 Log [Compound] nM
op-delta nor-BNI 120 Antagonist Naltrindole Naltrindole 100 2+ Measurement: Intracellular Ca 80 60 Detection method: FLIPR Fluorimetry 40 20 Incubation 25℃ (%) Inhibition 0 Stimulant DPDPE: 10 nM -20 -2 -1 0 1 2 3 4 5 Antagonist effect IC50: 1.62 nM Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]DADLE op-delta 125 Ligand concentration 0.5 nM Naltrindole 100 DPDPE Kd (nM) 0.7 nM 75 (± )trans-U-50488 Non specific naltrindole (1 µM) 50 25 Incubation 60 min/RT Inhibition % 0
Detection method Scintillation counting -25 -4 -3 -2 -1 0 1 2 3 4 5 Reference: Naltrindole(IC50:0.2nM) Log [Compound] nM
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op-kappa(κ-opioidReceptor)
Cell Line Name: HEK293/Kappa/Gα15 Species/Source: Human/Recombinant Gene Synonyms: OPRK1; KOR; OPRK; KOR-1; K-OR-1 Accession Number: NM_000912 G coupling Gi
Functional Assay-Calcium Flux Assay
op-kappa U69593 120 U-50488 Agonist U69593 DPDPE 100 2+ DAMGO Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 40
Incubation 25℃ (%) Activity 20 0 Agonist effect EC50: 11.9nM -20 -1 0 1 2 3 4 5 log [Compound] nM
nor-Binaltorphimine dihydrochl op-kappa Antagonist nor- naloxone Binaltorphiminedihydrochlo 120 5'-GNTI 100 ride 80 Measurement: Intracellular Ca2+ 60 40 20 Detection method: FLIPR Fluorimetry (%) Inhibition Incubation 25℃ 0 -20 -4 -2 0 2 4 6 Stimulant U69593: 20 nM Log [Compound] nM Antagonist effect IC50: 0.60nM
Radio ligand binding Assay
Ligand [3H]Diprenorphine op-kappa 125 DPDPE Ligand concentration 0.3 nM 100 (± )trans-U-50488 U69593 Kd (nM) 0.14 nM 75 50 Non specific trans-U-50488 (5 µM)
% Inhibition % 25
Incubation 60 min/RT 0
Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM Reference: U-50488 (IC50:41.8nM)
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op-mu (Μ-opioid Receptor)
Cell Line Name: HEK293/Mu/Gα15 Species/Source: Human/Recombinant Gene Synonyms: OPRM1; MOP; MOR; LMOR; MOR1; OPRM; M-OR-1 Accession Number: NM_000914 G coupling Gi
Functional Assay-Calcium Flux Assay
op-mu DAMGO Agonist DAMGO 120 DPDPE Measurement: Intracellular Ca2+ 100 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 (%) Activity 20 Agonist effect EC50: 3.6 nM 0 -20 -4 -2 0 2 4 6 Log [Compound] nM
op-mu CTOP Antagonist CTOP 120 Naltrindole hydrochloride Measurement: Intracellular Ca2+ 100 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 20 Inhibition (%) Inhibition Stimulant DAMGO: 20 nM 0 -20 Antagonist effect IC50: 62.3 nM -1 0 1 2 3 4 5 Log [Compound] nM
Radio ligand binding Assay
Ligand [3H]DAMGO op-mu 125 CTOP Ligand concentration 0.5 nM 100 Naltrindole DPDPE 75 Kd (nM) 0.3 nM DAMGO 50 Non specific naltrindole (1 µM)
% Inhibition % 25
Incubation 60 min/RT 0
Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM Reference: DAMGO(IC50:1.7 nM)
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V1A (Arginine Vasopressin Receptor 1A)
Cell Line Name: CHO/V1A Species/Source: Human/Recombinant Gene Synonyms: AVPR1A; V1aR; AVPR1; AVPR; V1a Accession Number: NM_000706 G coupling Gq
Functional Assay-Calcium Flux Assay
V1A AVP Agonist [Arg8]Vasopressin acetate 120 Oxytocin Measurement: Intracellular Ca2+ 100 80 Detection method: FLIPR Fluorimetry 60
Incubation 25℃ 40 (%) Activity 20 Agonist effect EC50: 0.0034 nM 0 -20 -4 -3 -2 -1 0 1 2 3 4 Log [Compound] nM
Antagonist [β-Mercapto-β,β-
cyclopentamethylenepropi
onyl1, O-Et-Tyr2, Val4, V1A 120 [β -Mercapto-β β -cyclopentame Arg8]-Vasopressin 100 [d(CH2)5Tyr(Me)2]-AVP 80 Measurement: Intracellular Ca2+ 60 Detection method: FLIPR Fluorimetry 40 20 (%) Inhibition Incubation 25℃ 0 Stimulant [Arg8]Vasopressin acetate: -20 -2 -1 0 1 2 3 4 0.12 nM Log [Compound] nM
Antagonist effect IC50: 6.551 nM
Radio ligand binding Assay
V1A Ligand [3H]AVP 125 AVP Ligand concentration 0.3 nM 100 [d(CH2)5Tyr(Me)2]-AVP Oxytocin Kd (nM) 0.26 nM 75 Non specific AVP (1 µM) 50
Incubation 60 min/RT Inhibition % 25 Detection method Scintillation counting 0 Reference: [d(CH2)5,Tyr(Me)2]-AVP -4 -3 -2 -1 0 1 2 3 4 Log [Compound] nM (IC50:0.92 nM)
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hCav1.2 (L-type calcium channel)
Channel Name: Cav1.2 Host Cells: T-Rex-293 Species/Source: Human/Recombinant Gene Synonyms: Cav1.2/beta2/ alpha2/delta Accession Number: NM_000719.6/ NM_000724.3/ NM_000722.2 Vector/Antibiotic: pcDNA4TO/Zeocin
Functional Assay- Manual Patch Clamp
Antagonist Nifedipine
Measurement: Current under voltage patch clamp
Detection method: Manual patch clamp
Temperature 25℃
Antagonist potency IC50: 2.85 nM
Inhibition of nifedipine on Cav1.2
1.2
1.0 IC50=2.85 nM 0.8 0.6 0.4 0.2
Normalized Current Normalized 0.0
10 -10 10 -9 10 -8 10 -7 log[Nifedipine](M)
Left: Raw traces of Cav1.2 current to show IV relationship Right: Nifedine inhibited Cav1.2 currents in dose manner with IC50 of 2.85 Nm
Ca2+-L (Calcium channel, voltage-dependent, L type)
Species/Source: rat cerebral cortex
Radio ligand binding Assay
2+ Ca -L Ligand [3H]-nitredipine 125 nitrendipine Ligand concentration 0.2 nM 100 nimodipine
Kd (nM) 0.27 nM 75 Non specific Nitrendipine (1 μM) 50
Incubation 60 min/RT Inhibition % 25 0 Detection method Scintillation counting
Reference: Nitrendipine (IC50:0.29 -4 -3 -2 -1 0 1 2 3 Log [Compound] nM nM)
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hERG channel (potassium channel)
Channel Name: hERG Host Cells: CHO Species/Source: Human/Recombinant Gene Synonyms: KCNH2 Accession Number: Swissprot Q12809 Vector/Antibiotic: pcDNA5/Hygromycin
Functional Assay- Manual Patch Clamp
Antagonist Amitriptyline; Terfenadine Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency IC50: 3.27 μM for amitriptyline and 9.38 for terfenadine
Sample traces of hERG currents in the presence of different concentrations of lidocaine
Inhibition of amitriptyline on hERG Inhibition of terfenadine on hERG
1.0 1.0
0.8 IC50=3.27 M 0.8 IC =9.38M 50 0.6 0.6
0.4 0.4
0.2 0.2
Normalized current Normalized Normalized Current Normalized 0.0 0.0 10 -7 10 -6 10 -5 10 -4 10 -9.0 10 -8.5 10 -8.0 10 -7.5 10 -7.0 10 -6.5 log[Amitriptyline](M) log[Terfenadine](M)
Dose response curve of amitriptyline and terfenadine on hERG channels
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hERG (potassium channel)
Cell Line Name: Hela Species/Source: Human Gene Synonyms: KCNH2 Accession Number: Swissprot Q12809
Radio ligand binding Assay
hERG Ligand [3H]- dofetilite 125 Ligand concentration 1 nM dofetilite 100 Kd (nM) 1.3 nM 75 Non specific dofetilite (1 μM) 50 Incubation 60 min/RT
% Inhibition % 25
Detection method Scintillation counting 0
Reference: dofetilite(IC50:5.15 nM) -2 -1 0 1 2 3 Log [Compound] nM
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hKv1.5 (Potassium channel)
Channel Name: hKv1.5 Host Cells: CHO Species/Source: Human/Recombinant Gene Synonyms: KCNA5 Accession Number: NM_002234.2
Functional Assay- Manual Patch Clamp
Antagonist 4-AP Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency IC50: 217 μM
IV curve of Kv1.5 1.0
0.8
0.6
0.4
V1/2 =1.7 0.4mV 0.2 Current Normalized 0.0 -60 -40 -20 0 20 40 60 Voltage / mV
Sample traces of Kv1.5 current (left) and its IV curve (right)
Inhibition of 4-AP on Kv1.5
1.0
0.8 IC50=217M 0.6
0.4 0.2
Normalized Current Normalized 0.0 10 -5 10 -4 10 -3 log[4-AP]M
Dose response curve of 4-AP on Kv1.5 channels
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Kv(Potassium channel)
Species/Source: rat cerebral cortex
Radio ligand binding Assay
Kv 125 Ligand [125I] α-DENDROTOXIN charybdotoxin Ligand concentration 0.01 nM 100 ML277 Kd (nM) 0.04 nM 75 Non specific charybdotoxin (1μM) 50
% Inhibition % 25 Incubation 30 min/RT 0 Detection method Scintillation counting -4 -2 0 2 4 Reference: charybdotoxin (IC50:3.9 Log [Compound] nM nM)
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hKCNQ1 (Potassium channel)
Channel Name: hKCNQ1+ MinK Host Cells: T-REx-CHO Species/Source: Human/Recombinant Gene Synonyms: KvLQT1/minK Accession Number: NM_000218/ NM_000219 Vector/Antibiotic: pcDNA4TO/Zeocin, pcDNA3.1/Geneticin
Functional Assay- Manual Patch Clamp
Antagonist Chromanol 293B Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency IC50: 8.17 μM
Sample traces of KCNQ1 current (left) and its IV curve (right)
Inhibition of chromanol 293B on KCNQ1 1.0 0.8 IC50=8.17M 0.6
0.4
0.2
Normalized Current Normalized 0.0 10 -6 10 -5 10 -4 log[Chromanol 293B](M)
Dose response curve of Chromanol 293B on KCNQ1 channels
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hNav1.5 (Sodium channel)
Channel Name: Nav1.5 Host Cells: HEK 293 Species/Source: Human/Recombinant Gene Synonyms: SCN5A Accession Number: NM_000335.4 Vector/Antibiotic: pcDNA3.1/Geneticin
Functional Assay- Manual Patch Clamp
Antagonist TTX， tetrecane
Measurement: Current under voltage patch clamp
Detection method: Manual patch clamp
Temperature 25℃
Antagonist potency IC50: 1.62 μM for TTX, 428.5μM for lidocaine
Time (ms) Time (s) 22 24 26 28 30 200 400 600 0 0 3 mM 1 mM 3 mM 1 mM -2000 -2000 300 M 300 M 100 M -4000 Control 30 M Current (pA) -4000 100 M Current (pA) Control 30 M -6000 -6000 Control
Left: Sample Nav1.5 current traces in the presence of lidocaine at different concentrations.
Right: IT plot of Nav1.5 peak current in the presence or absence of lidocaine.
Inhibition of TTX on Nav1.5 The inihibition of lidocaine on Nav1.5
1.0 1.0
0.8 IC50=428. 5M IC50=1.62 M 0.6 0.5 0.4
Normalized Current Normalized 0.2
Normalized Current Normalized 0.0 10 -8 10 -7 10 -6 10 -5 10 -4 0.0 10 -5 10 -4 10 -3 10 -2 log[TTX](M) log[Lidocaine](M)
Dose response curve of TTX and lidocaine on Nav1.5 channels
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5-HT3A (Serotonin receptor)
Channel Name: 5-HT3A Host Cells: BacMam system (HEK) Species/Source: Human/Recombinant Gene Synonyms: HTR3A Accession Number: BC145743 Vector/Antibiotic: pFastBac (Bac-to-Bac Baculovirus Expression System)
Functional Assay- Manual Patch Clamp
Antagonist 5-HT Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency EC50: 1.70 μM
A 0.1 uM 1 uM 3 uM 10 uM 30 uM 100 uM 5-HT 5-HT 5-HT 5-HT 5-HT 5-HT
1 nA 10 s
B C
Sample traces and dose response curve of 5-HT induced current in 5-HT3A receptor. The interval of 5-HT application is 3 min. A, Sample traces. All traces were from same cell. B, the sample traces in A were superimposed. C,Dose response curve of 5-HT. The EC50 is 1.7 uM.
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5HT3A (Serotonin receptor)
Cell Line Name: BacMam system (HEK)/5-HT3A Species/Source: Human/Recombinant Gene Synonyms: HTR3A Accession Number: BC145743
Radio ligand binding Assay
5HT Ligand [3H]BRL 43694 3A 125 Ligand concentration 0.5 nM MDL72222 100 Serotonin 8-OH-DPAT Kd (nM) 0.13 nM 75
Non specific MDL 72222 (10 μM) 50
Incubation 60 min/RT Inhibition % 25
Detection method Scintillation counting 0
Reference: MDL 72222 (IC50:22.1 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM nM)
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nAChRα7 (Nicotinic receptor)
Channel Name: nAChRα7 Host Cells: T-Rex-293 Species/Source: Human/Recombinant Gene Synonyms: CHRNA7 and RIC3 Accession Number: NP_000737.1/ NP_001193600.1 Vector/Antibiotic: PcDNA4TO/Zeocin, pcDNA3.1/Geneticin
Functional Assay- Manual Patch Clamp
Antagonist ACh Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency EC50: 160 μM
ACh ACh ACh ACh ACh 100 μM 300 μM 1000 μM 3000 μM 10000 μM
300 300
pA 100 ms
The up pannel is the sample traces of ACh induced currents. All the currents were from same cell. The EC50 of ACh induced peak current is 160 uM. The peak current area was also calculated. All the peak current values and current area were normalized to the one induced by 1mM ACh. Data were presented as Mean ±SD (n=4)
PNU+ 300 μM ACh
0 μM 300 μM ACh 0.1 μM 0.3 μM
300 300 1 μM pA
1500 1500 3 μM 200ms 10 μM
pA 0 μM 5 s
PNU-120596 enhance ACh-induced currents in dose response manner. PNU dose response curve in the presence of 300 μM of Ach. All the currents were normalized to the one induced by 10 uM PNU plus 300 uM ACh.Data were presented as Mean ±SD (n=5)
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nAChR-alpha7 (Nicotinic Acetylcholine receptor)
Cell Line Name: T-Rex-293/nAChR-alpha7 Species/Source: Human/Recombinant Gene Synonyms: CHRNA7 and RIC3 Accession Number: NP_000737.1/ NP_001193600.1
Radio ligand binding Assay
nAChR-alpha7 Ligand [3H]-Methyllycaconitine 125 Ligand concentration 1 nM trifluridine 100 methyllycaconitine Kd (nM) 2.8 nM 75 Non specific Trifluridine (10 μM) 50 Incubation 60 min/RT
% Inhibition % 25
Detection method Scintillation counting 0
Reference: Methyllycaconitine -4 -3 -2 -1 0 1 2 3 4 5 (IC50:6.9 nM) Log [Compound] nM
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GABAA receptor ( α1β3γ2)
Channel Name: GABAA receptor Host Cells: HEK293 Species/Source: Human/Recombinant Gene Synonyms: GABRA1/GABRB3/GABRG2 Accession Number: NM_000806/ NM_000814/ NM_000816 Vector/Antibiotic: pcDNA3.1/ Geneticin
Functional Assay- Manual Patch Clamp
Antagonist GABA Measurement: Current under voltage patch clamp Detection method: Manual patch clamp Temperature 25℃ Antagonist potency
GAB GABA
A
GABA GABA+DZP
Samples traces from Qpatch to show GABA induced current in GABAA receptor ( α1β3γ2) in the presence and absence of DZP.
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GABAA (GABAA -α1β3γ2 receptor)
Cell Line Name: HEK293/GABAA Species/Source: Human/Recombinant Gene Synonyms: GABRA1/GABRB3/GABRG2 Accession Number: NM_000806/ NM_000814/ NM_000816
Radio ligand binding Assay
GABAA Ligand [3H]flunitrazepam 125 Ligand concentration 0.4 nM Dizepam 100 Kd (nM) 2.7 nM Flumazenil 75 Non specific Flumazenil (10 μM) 50 Incubation 60 min/4°C Inhibition % 25 Detection method Scintillation counting 0 Reference: Flumazenil(IC50:3.4 -3 -2 -1 0 1 2 3 4 5 nM) Log [Compound] nM
NMDA (N-methyl-D-aspartate receptor)
Species/Source: rat cerebral cortex
Radio ligand binding Assay
NMDA
Ligand [3H]CGP 39653 125 L-Glutamate Ligand concentration 5 nM 100 CGS19755
Kd (nM) 13.5 nM 75 Non specific L-Glutamate (100 μM) 50 25
Incubation 30 min/RT Inhibition %
Detection method Scintillation counting 0
Reference: CGS 19755 (IC50:156.3 -25 0 1 2 3 4 5 nM) Log [Compound] nM
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DAT (Dopamine Transporter)
Cell Line Name: HEK293/DAT Species/Source: Human/Recombinant Gene Synonyms: SLC6A3 Accession Number: NM_001044
Functional Assay-Transporter uptake Assay
DAT BTCP 125 Reference BTCP GBR12909 100 nomifensine Dopamine Measurement: Dye uptake 75 Detection method: Fluorescence 50 25 Reference effect: BTCP(IC50:75 nM) inhibition % 0
-25 -2 0 2 4 6 Log [Compound] nM
Radio ligand binding Assay
DAT Ligand [3H]-WIN 35,428 125 BTCP GBR 12909 Ligand concentration 12.5 nM 100 Nomifensine
Kd (nM) 18 nM 75
Non specific BTCP (10 μM) 50 25 Incubation 120min/4°C inhibition % 0 Detection method Scintillation counting -25 Reference: BTCP(IC50:9.5 nM) -2 -1 0 1 2 3 4 Log [Compound] nM
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5HTT (5-HT Transporter)
Cell Line Name: HEK293/5-HTT Species/Source: Human/Recombinant Gene Synonyms: SLC6A4 Accession Number: NM_001045
Functional Assay-Transporter uptake Assay
5HTT 120 Reference Citalopram Imipramine 100 Serotonin Measurement: Dye uptake 80 Citalopram Zimetidine Detection method: Fluorescence 60 40 Reference effect: Citalopram(IC50:8.4 nM) inhibition % 20
0 -2 0 2 4 6 Log[compound] nM
Radio ligand binding Assay
5HTT Ligand [3H]imipramine 125 Imipramine Ligand concentration 2 nM 100 Serotonin Kd (nM) 2.4 nM 75 50 Non specific imipramine (10 μM)
% Inhibition % 25
Incubation 60 min/RT 0
Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM Reference imipramine(IC50:5.1nM)
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NET (Noradrenaline Transporter)
Cell Line Name: HEK293/NET Species/Source: Human/Recombinant Gene Synonyms: SLC6A2 Accession Number: NM_001043
Functional Assay-Transporter uptake Assay
Reference Nisoxetine NET Nisoxetine hydrobromide 125 Measurement: Dye uptake GBR12909 100 Imipramine Protriptyline Detection method: Fluorescence 75 Reference effect: Nisoxetine (IC50:32 nM) 50 25 inhibition % 0
-25 -2 0 2 4 6 Log[compound] nM
Radio ligand binding Assay
Ligand [3H]nisoxetine NET 125 Ligand concentration 1 nM Protriptyline 100 imipramine
Kd (nM) 6.9 nM 75 Non specific Protriptyline (10 μM) 50
Incubation 120min/4°C Inhibition % 25 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference Protriptyline(IC50:3.2 Log [Compound] nM nM)
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Androgen receptor
Cell Line Name: GeneBLAzer ® AR-UAS-bla GripTite™ Cells Species/Source: ligand-binding domain of the rat AR
Functional Cellular assay
AR Agonist Testosterone 120 Testosterone Measurement: CCF-4 100 progesterone Detection method: FRET 80 17-beta-estradiol 60 Incubation 25℃ 40
Agonist effect EC50: 1.9 nM (%) Activity 20 0 0 2 4 6 -20 Log [Compound] nM
Antagonist Cyproterone Acetate AR 120 Cyproterone acetate Measurement: CCF-4 100 ARN-509 Detection method: FRET 80 60 Incubation 25℃ 40 20 Stimulant Testosterone : 1.89 nM (%) Inhibition 0 0 2 4 6 Antagonist effect IC50: 55.8 nM -20 Log [Compound] nM
Androgen receptor
Cell Line Name: LNCap Species/Source: Human/Cytosol Gene Synonyms: AR Accession Number: NM_000044 and NM_001011645
Radio ligand binding Assay
AR Ligand [3H]methyltrienolone 125 Ligand concentration 1 nM 17-beta-estradiol 100 progesterone Kd (nM) 0.33 nM 75 Non specific Progesterone (10 μM) 50
Incubation 24hr/4°C
% Inhibition % 25
Detection method Scintillation counting 0
Reference: Progesterone (IC50:8.42 -3 -2 -1 0 1 2 3 4 5 nM) Log [Compound] nM
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Glucocorticoid receptor
Cell Line Name: GeneBLAzer ® GR-UAS-bla HEK 293T cells Species/Source: Human GR ligand-binding domain
Functional cellular assay
GR Dexamethasone Agonist Dexamethasone 125 triamcinolone Measurement: CCF-4 100
Detection method: FRET 75 Incubation 25℃ 50
Agonist effect EC50: 2.2 nM % Activity 25 0
-25 -4 -2 0 2 4 Log [Compound] nM
GR Antagonist Mifepristone 150
Measurement: CCF-4 Mifepristone Ketoconazole Detection method: FRET 100 Incubation 25℃ 50
Inhibition (%) Inhibition Stimulant Dexamethasone : 4.06 nM
0 Antagonist effect IC50: 0.85 nM -2 0 2 4 6 Log [Compound] nM
Glucocorticoid receptor
Cell Line Name: IM-9 Species/Source: Human/Cytosol Gene Synonyms: NR3C1 Accession Number: NM_000176
Radio ligand binding assay
GR
Ligand [3H]dexamethasone 125 dexamethasone Ligand concentration 1.5 nM 100 triamcinolone progesterone 75 Kd (nM) 3.23 nM 17-beta-estradiol
Non specific triamcinolone(10 μM) 50
Incubation 18hr/4°C Inhibition % 25 0 Detection method Scintillation counting
Reference: dexamethasone(IC50: -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM 3.3nM)
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LCK
Enzyme: Human recombinant (insect cells) LCK Gene Synonyms: LSK; YT16; IMD22; p56lck; pp58lck 120 stausporine 100 856-1-3 Accession Number: NP_005347 80 pp1 Substrate: Z’-LYTE®Tyr2 Peptide (2μM) 60 pp2 40 20 Measured Product: Cleaved and uncleaved peptides (%) Inhibition Detection Method: FRET 0 -20 -2 0 2 4 6 Reference: Stausporine (IC50: 3.02nM) Concentration (nM)
PDE3A
Enzyme Source: Human recombinant (sf-9) PDE3A Trequinsin Gene Synonyms: HTNB;CGI-PDE;CGI-PDEA; CGI-PDE-A 120 Milrinone 100 Cilostamide Accession Number: NC_000012.12 80 Substrate: cAMP (20 nM) 60 40 20 Detection Method: HTRF (%) Inhibition 0 Reference: Milrinone (IC50: 411nM) -20 -4 -2 0 2 4 6 Log[compound] nM
PDE4D2
Enzyme: Human recombinant (sf-9) PDE4D2 Gene Synonyms: DPDE3; PDE43; STRK1; ACRDYS2; 140 R020-1724 120 IBMX HSPDE4D; PDE4DN2 100 rolipram 80 Accession Number: NC_ 000005.10 60 Substrate: cAMP (20 nM) 40 Inhibition (%) Inhibition 20 Detection Method: HTRF 0 -20 -2 0 2 4 6 8 Reference: IBMX (IC50: 31uM) Log [Compound] nM
Enzyme: Human recombinant (sf-21) COX-1 Gene Synonyms: PTGS1; COX3; PHS1; PCOX1; PES-1; Diclofenac sodium salt 150 NS-398 PGHS1; PTGHS; PGG/HS; PGHS-1 SC-560 100 indomethacin Accession Number: NC_000009.12 50 Substrate: Arachidonic acid (3μM)
Measured product: Resorufin (%) Inhibition 0
Detection Method: Fluorescence -50 -2 0 2 4 6 Reference: Diclofenac sodium salt (IC50: Log [Compound] nM 60.3nM)
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COX-2
Enzyme: Human recombinant (sf-21) COX-2 Gene Synonyms: PTGS2; COX-2; PHS-2; PGG/HS; NS-398 125 Diclofenac sodium salt PGHS-2; hCox-2; GRIPGHS 100 indomethacin rofecoxib Accession Number: NC_000001.11 75 50 Substrate: Arachidonic acid (3μM) 25
Inhibition (%) Inhibition Measured product: Resorufin 0 -25 Detection Method: Fluorescence -2 0 2 4 6 Log [Compound] nM Reference: NS-398 (IC50: 224nM)
Acetylcholinesterase
Enzyme: Human recombinant (HEK-293) Acetylcholinesterase Galanthamine Neostigmine bromide Gene Synonyms: YT; ACEE; ARACHE; N-ACHE 120 Tacrine hydrochloride Accession Number: NC_000007.14 100 80 Substrate: Acetylthiocholine iodide (200 μM) 60 40 Measured product: Thio-conjugate 20
Inhibition (%) Inhibition Detection Method: Photometry 0 -20 Reference: -2 0 2 4 6 Galantamine (IC50: 2.1μM) Log [Compound] nM
MAO-A
Enzyme: Human recombinant (BTI) MAO-A Clorgyline 120 Gene Synonyms: BRNRS; MAOA R-(-)-Deprenyl 100 Ro 41-1049 Accession Number: NC_000023.11 80 Substrate: luciferin derivative (8 μM) 60 40 20 Measured product: Methyl ester luciferin (%) Inhibition 0 Detection Method: Luminescent -20 -2 0 2 4 6 Reference: Clorgyline (IC50: 3.1nM) Log[Compound] (nM)
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