鱼藤酮诱导BV2小胶质细胞半胱氨酰白三烯受体1表达变化

Q421; 目的:制备和鉴定半胱氨酰白三烯受体1(CysLT1受体)抗体,以此观察鱼藤酮诱导BV2小胶质细胞损伤过程中CysLT1受体表达变化.方珐:以KLH偶联的CysLT1受体多肽(小鼠)免疫家兔制备多克隆抗体,用ELISA法测定抗体效价,以抗原阻断法测定抗体敏感性及特异性.用新制备的抗体以Western blotting检测鱼藤酮(0.01～1 μmoL/L,作用24 h)损伤BV2细胞过程中,BV2细胞上CysLT1受体的表达变化,并以RT-PCR检测验证CysLT1受体mRNA表达水平,同时以免疫组化观察CysLT1受体的亚细胞分布变化.结果:ELISA测定显示,制备的兔抗小鼠Cys...

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Published in浙江大学学报（医学版） Vol. 40; no. 2; pp. 131 - 138
Main Authors 骆江云, 张壮, 余舒莹, 赵冰, 赵春贞, 王欣欣, 方三华, 张纬萍, 张丽慧, 魏尔清, 卢韵碧
Format Journal Article
LanguageChinese
Published 浙江大学医学院药理学系,浙江杭州,310058%杭州师范大学医学神经生物学市级重点实验室、基础医学部药理学教研室,浙江杭州,310036 2011
Subjects
抗体
BV2小胶质细胞
半胱氨酸
受体,白三烯
半胱氨酰白三烯受体
鱼藤酮/药理学
多克隆抗体
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ISSN1008-9292
DOI10.3785/j.issn.1008-9292.2011.02.003

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Abstract Q421; 目的:制备和鉴定半胱氨酰白三烯受体1(CysLT1受体)抗体,以此观察鱼藤酮诱导BV2小胶质细胞损伤过程中CysLT1受体表达变化.方珐:以KLH偶联的CysLT1受体多肽(小鼠)免疫家兔制备多克隆抗体,用ELISA法测定抗体效价,以抗原阻断法测定抗体敏感性及特异性.用新制备的抗体以Western blotting检测鱼藤酮(0.01～1 μmoL/L,作用24 h)损伤BV2细胞过程中,BV2细胞上CysLT1受体的表达变化,并以RT-PCR检测验证CysLT1受体mRNA表达水平,同时以免疫组化观察CysLT1受体的亚细胞分布变化.结果:ELISA测定显示,制备的兔抗小鼠CysLT1受体抗体的效价大于1/32728,对CysLT1受体的特异性强,对CysLT2受体和GPR17基本无交叉反应.Western blotting和RT-PCR显示,鱼藤酮作用BV2细胞24 h剂量依赖性诱导CysLT1受体mRNA及蛋白水平表达升高;免疫组化显示在鱼藤酮诱导BV2细胞损伤过程中,随鱼藤酮荆量增加,CysLT1受体发生核移位.结论:制备的CysLT1受体多克隆抗体具有高效价和高特异性,能满足Western blotting和免疫组化检测的要求;CysLT1受体参与鱼藤酮诱导的BV2细胞损伤.
AbstractList Q421; 目的:制备和鉴定半胱氨酰白三烯受体1(CysLT1受体)抗体,以此观察鱼藤酮诱导BV2小胶质细胞损伤过程中CysLT1受体表达变化.方珐:以KLH偶联的CysLT1受体多肽(小鼠)免疫家兔制备多克隆抗体,用ELISA法测定抗体效价,以抗原阻断法测定抗体敏感性及特异性.用新制备的抗体以Western blotting检测鱼藤酮(0.01～1 μmoL/L,作用24 h)损伤BV2细胞过程中,BV2细胞上CysLT1受体的表达变化,并以RT-PCR检测验证CysLT1受体mRNA表达水平,同时以免疫组化观察CysLT1受体的亚细胞分布变化.结果:ELISA测定显示,制备的兔抗小鼠CysLT1受体抗体的效价大于1/32728,对CysLT1受体的特异性强,对CysLT2受体和GPR17基本无交叉反应.Western blotting和RT-PCR显示,鱼藤酮作用BV2细胞24 h剂量依赖性诱导CysLT1受体mRNA及蛋白水平表达升高;免疫组化显示在鱼藤酮诱导BV2细胞损伤过程中,随鱼藤酮荆量增加,CysLT1受体发生核移位.结论:制备的CysLT1受体多克隆抗体具有高效价和高特异性,能满足Western blotting和免疫组化检测的要求;CysLT1受体参与鱼藤酮诱导的BV2细胞损伤.
Author 赵春贞
魏尔清
王欣欣
张丽慧
张壮
张纬萍
骆江云
方三华
余舒莹
赵冰
卢韵碧
AuthorAffiliation 浙江大学医学院药理学系,浙江杭州,310058%杭州师范大学医学神经生物学市级重点实验室、基础医学部药理学教研室,浙江杭州,310036
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Author_FL ZHAO Chun-zhen
ZHANG Zhuang
LU Yun-bi
WANG Xin-xin
ZHANG Wei-ping
ZHAO Bing
ZHANG Li-hui
LUO Jiang-yun
YU Shu-ying
FANG San-hua
WEI Er-qing
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Keywords 抗体
BV2小胶质细胞
半胱氨酸
受体,白三烯
半胱氨酰白三烯受体
鱼藤酮/药理学
多克隆抗体
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PublicationTitle 浙江大学学报（医学版）
PublicationTitle_FL JOURNAL OF ZHEJIANG UNIVERSITY(MEDICAL SCIENCES)
PublicationYear 2011
Publisher 浙江大学医学院药理学系,浙江杭州,310058%杭州师范大学医学神经生物学市级重点实验室、基础医学部药理学教研室,浙江杭州,310036
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Snippet Q421; 目的:制备和鉴定半胱氨酰白三烯受体1(CysLT1受体)抗体,以此观察鱼藤酮诱导BV2小胶质细胞损伤过程中CysLT1受体表达变化.方珐:以KLH偶联的CysLT1受体多肽(小鼠)免疫家兔制备多克隆抗体,用ELISA法测定抗体效价,以抗原阻断法测定抗体敏感性及特异性.用新制备的抗体以Western...
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StartPage 131
Title 鱼藤酮诱导BV2小胶质细胞半胱氨酰白三烯受体1表达变化
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