Application of furanocoumarins in the preparation of antiallergic drugs
A technology of furanocoumarins and compounds, applied in the field of biomedicine, can solve problems such as painful life-threatening, hypersensitivity reactions, and elevated calcium ion concentration in mast cells, and achieve the effect of inhibiting hypersensitivity reactions
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Embodiment 1
[0024] 1. Experimental materials
[0025] Instrument: Automatic microplate reader was purchased from Bio-Rad (California, USA).
[0026] Cell line: KU812 was cultured in IMEM medium with 10% serum, plus 1:100 penicillin-streptomycin. The medium was half-replaced with fresh medium every other day to maintain the cells at 2 × 10 6 density of cells per milliliter.
[0027] Main reagent: TM buffer solution (composition (g / L): 6.954 NaCl, 0.353 KCl, 2.383 HEPES, 0.162 KH 2 PO 4 , 0.282 CaCl 2 , 0.143 MgSO 4 , 0.991 glucose, 1 bovine serum albumin. Dimethyl sulfoxide was purchased from Guangzhou Jinhuada Chemical Reagent Co., Ltd., and C48 / 80 was purchased from Sigma-Aldrich (St.Louis, MO, USA). The imperatorin, isoimperatorin, and pectinoids were all purchased from Chengdu Pufeid Biotechnology Co., Ltd. (Chengdu, China). Triton X-100 was purchased from Kehao Bioengineering Company (Xi'an, China), D-glucose Tianjin Kemiou Chemical Reagent Company (Tianjin, China), bovine ser...
Embodiment 2
[0052] 1. Experimental materials
[0053] Instrument: LC-2010AHT (SHIMAZU), self-made column packing machine.
[0054] Cell line: MrgprX2-HEK293 cells were cultured in DMEM medium with 10% fetal bovine serum, plus 1:100 penicillin-streptomycin in 5% CO 2 , in an environment of 37°C. The medium was replaced with fresh medium every other day to keep the cells healthy.
[0055] Main reagents: DMEM medium and fetal bovine serum were purchased from Hyclone (Logan, Utah, USA), and macroporous silica gel was purchased from Qingdao Meigao Group Co., Ltd. (Qingdao, China).
[0056] 2. Experimental method
[0057] 2.1 Preparation of MrgprX2-HEK293 cell membrane stationary phase
[0058] Take the above-mentioned MrgprX2-HEK293 cells (about 7×10 6 cells) suspension, centrifuge at 1000rpm, 4°C for 5min, discard the supernatant, and take the bottom cells; wash the centrifuged cells twice with 5mL saline, and then add Tris-HCl 5mL to the obtained cells Ultrasound for 30 min in an ice b...
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