20160519-中国互联网协会-中国移动互联网发展状况及其安全报告(2016)(简版).pdf
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1、ClemizoleCat. No.: HY-30234CAS No.: 442-52-4分式: CHClN分量: 325.84作靶点: Histamine Receptor作通路: GPCR/G Protein; Immunology/Inflammation储存式: Please store the product under the recommended conditions in the COA.BIOLOGICAL ACTIVITY物活性 Clemizole是 H1受体拮抗剂,还能抑制 NS4B的 RNA结合及 HCV的复制 。IC it is very rapidly biotra
2、nsformed into a glucuronide (M14) and a dealkylated metabolite (M12) and into a variety of lesser metabolites in C57BL/6J mice3.PROTOCOLHuh7.5 cells are maintained in DMEM supplemented with 1% L-glutamine, 1% penicillin, 1% streptomycin, 1 nonessential amino acids and 10% FBS. Cell lines are passage
3、d twice weekly after treatment with 0.05% trypsin-0.02% EDTA and seeding at a dilution of 1:5. Subconfluent Huh7.5 cells are trypsinized and collected by centrifugation at 700g for 5 min. The cells are then washed three times in ice-cold RNase-free PBS and resuspended at 1.5107 cells/mL in PBS. Wild
4、-type or mutant FL-J6/JFH-5C19Rluc2AUbi RNA for electroporation is generated by transcription of XbaI linearized DNA templates using the T7 MEGAscript kit, followed by purification (RNA transcription and fluorescent labeling). We mixed 5 g of RNA with 400 L of washed Huh7.5 cells in a 2-mm-gap cuvet
5、te (BTX) and immediately pulsed (0.82 kV, five 99 s pulses) with a BTX-830 electroporator. After a 10 min recovery at 25C, pulsed cells are diluted into 10 mL of prewarmed growth medium. Cells from several electroporations are pooled to a common stock and seeded in 6-well plates (5105 cells per well
6、). After 24 h, Cell Assay 1Product Data Sheet InhibitorsAgonistsScreening Librarieswww.MedChemE1medium is replaced and cells are grown in the presence of serial dilutions of the various inhibitory compounds (e.g., Clemizole hydrochloride) identified in the screen. Seventeen commercially available co
7、mpounds, out of the 18 identified, are analyzed. Untreated cells are used as a negative control for water-soluble compounds. For compounds (e.g., Clemizole hydrochloride) solubilized in DMSO, untreated cells are grown in the presence of corresponding concentrations of the solvent as a negative contr
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