Recombinant human MAO-A and -B were purchased from Sigma. The inhibitory activity was determined using MAO-Glo assay kit (Promega). In brief, following the manufacturer’s protocol, assays were performed in 384-well white plates (Corning) using MAO-A or -B (100 nM) with a final volume of 20 μL. Reactions were quenched after 60 min by adding reconstituted luciferin detection reagent (20 μL/well); 20 min after addition of the detection reagent, the luminescence of each well was measured using Beckman DTX-880 microplate reader. IC50 calculation was similar to that of LSD1.
Mao a
Manufactured by Corning
MAO-A is a laboratory equipment product manufactured by Corning. It is a specialized device used for the measurement and analysis of monoamine oxidase A enzyme activity. The core function of MAO-A is to facilitate the quantification of this enzyme, which plays a role in the metabolism of various neurotransmitters. This device provides researchers and scientists with a tool to study the activity and dynamics of monoamine oxidase A in various experimental and research settings.
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Lab products found in correlation
Mao glo assay kit, by Promega (1 mentions)
White 384 well plate, by Corning (1 mentions)
Dtx880, by Beckman Coulter (1 mentions)
Nadph regenerating system solution b, by Corning (1 mentions)
Ultrapool human liver microsomes, by Corning (1 mentions)
Ugt1a10, by Corning (1 mentions)
Nadph regenerating system solution a, by Corning (1 mentions)
Ugt reaction mix solution a, by Corning (1 mentions)
Ugt reaction mix solution b, by Corning (1 mentions)
Midazolam, by Fujifilm (1 mentions)
Phenylephrine, by Fujifilm (1 mentions)
Eletriptan, by Merck Group (1 mentions)
Sumatriptan, by Tokyo Chemical Industry (1 mentions)
Sertraline, by Fujifilm (1 mentions)
Fatty acid free human serum albumin, by Merck Group (1 mentions)
3 protocols using mao a
1
Measuring MAO-A and -B Inhibitory Activity
2
Comprehensive In Vitro ADME Toolkit
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Corning UltraPool human liver microsomes (HLM) 150 (20 mg/mL), human intestinal microsomes (HIM) pool (10 mg/mL) as well as recombinant human CYP enzymes (with P450 oxidoreductase, OR) including CYP1A2+OR (0.5 nmol), CYP2B6+OR (0.5 nmol), CYP2C8+OR (1.0 nmol), CYP2C9*1+OR (1 nmol), CYP2C19+OR (0.5 nmol), CYP2D6*1+OR (0.5 nmol), CYP2E1+OR + cytochrome b5 (1 nmol), CYP3A4+OR + cytochrome b5 (0.5 nmol), MAO-A (5 mg/mL), MAO-B (5 mg/mL), and UGT1A10 (5 mg/mL) were purchased from Corning Life Sciences B.V (Amsterdam, Netherlands). All microsomes and recombinant enzyme solutions were aliquoted and stored at −80 °C until further use. Co-factors including NADPH regenerating system solution A (25 mM NADP+, 66 mM glucose-6-phosphate, and 66 mM MgCl2 in water), NADPH regenerating system solution B (40 U/mL glucose-6-phosphate dehydrogenase in 5 mM sodium citrate), UGT reaction mix solution A (25 mM uridine 5′-diphospho-glucuronic acid in water), and UGT reaction mix solution B (250 mM Tris-HCl, 40 mM MgCl2, and 0.125 mg/mL alamethicin in water) were also obtained from Corning Life Sciences B.V.
3
Liver Microsome Preparation and Characterization
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Midazolam, rizatriptan, phenylephrine and sertraline were purchased from Wako Pure Chemical Industries (Osaka, Japan). Sumatriptan and S-citalopram were purchased from Tokyo Chemical Industry (Tokyo, Japan). Imipramine was purchased from Nakalai Tesque (Kyoto, Japan). Eletriptan and fatty acid-free human serum albumin (HSA) were obtained from Sigma-Aldrich (St. Louis, MO). All other chemicals and reagents were analytical-grade products from commercial sources. Pooled HLMs (50 donors, #PLo50BA), CD-1 mouse liver microsomes, SD rat liver microsomes and beagle dog liver microsomes were purchased from Thermo Fisher Scientific (Waltham, MA). Pooled HLMt (5 donors, #1110152) was purchased from Sekisui XenoTech.
(Kansas City, KS). An MAO expression system (Supersomes: MAO-A, MAO-B and control) was purchased from Corning (Corning, NY). Individual HLMs and HLMt were prepared according to the previously reported method (13) . Human liver blocks were obtained from Human and Animal Bridging Research Organization (Chiba, Japan). The study protocol was approved by the ethical committees of the School of Medicine in Kanazawa University. Individual HLMs and HLMt were independently prepared to prevent loss derived from sequential purification.
(Kansas City, KS). An MAO expression system (Supersomes: MAO-A, MAO-B and control) was purchased from Corning (Corning, NY). Individual HLMs and HLMt were prepared according to the previously reported method (13) . Human liver blocks were obtained from Human and Animal Bridging Research Organization (Chiba, Japan). The study protocol was approved by the ethical committees of the School of Medicine in Kanazawa University. Individual HLMs and HLMt were independently prepared to prevent loss derived from sequential purification.
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