Method for screening compound with lipase inhibition activity by magnetic bead separation and application of method
A technology for inhibiting activity and separation of magnetic beads, used in material separation, analytical materials, measurement devices, etc.
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Embodiment 1
[0021] Example 1 Preparation of magnetic beads bound to lipase
[0022] 1. Reagents:
[0023] Lipase (extracted from porcine pancreatin), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), 2-(N-morpholine)ethanesulfonic acid (MES ), N-hydroxysuccinimide (NHS) were purchased from Simga Company; carboxyl magnetic beads (specification: 10 mg / ml) were purchased from Invitrogen Company; other reagents were of analytical grade.
[0024] Preparation of 25 mmol / L MES solution: Weigh an appropriate amount of MES, add water to dissolve, add 1 mol / L sodium hydroxide solution to adjust the pH to 6, and prepare a MES solution with a concentration of 25 mmol / L.
[0025] Preparation of 50 mg / ml EDC solution: Weigh an appropriate amount of EDC and quickly dissolve it in cold 25 mmol / L MES solution to prepare an EDC solution with a concentration of 50 mg / ml. Prepared fresh just before use.
[0026] Preparation of 50 mg / ml NHS solution: Weigh an appropriate amount of NHS and ...
Embodiment 2
[0036] Compound screening with lipase inhibitory activity in the total flavonoids extract of Fructus Aurantii in embodiment 2
[0037] 1. Drugs and reagents:
[0038] The standard products of naringin and 4-methylumbelliferone oleate were purchased from Sigma, the standard products of hesperidin and neohesperidin were purchased from Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd., and the standard products of citrusin were purchased from Shanghai Tongtian Biotechnology Co., Ltd., the total flavonoid extract of Aurantii Fructus was purchased from Changsha Kangerjia Pharmaceutical Group Co., Ltd.
[0039] Preparation of 10 mmol / L phosphate (PBS) buffer solution Weigh an appropriate amount of sodium dihydrogen phosphate, add water to dissolve, and make a solution of sodium dihydrogen phosphate with a concentration of 10 mmol / L, and weigh an appropriate amount of disodium hydrogen phosphate, add water to dissolve, Prepare a disodium hydrogen phosphate solution with ...
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