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大鼠淋巴细胞培养上清液总IgG 含量ELISA 检测方法的建立
引用本文:单纯筱王宇尹磊淼徐玉东魏颖冉君刘晓燕刘奇杨永清.大鼠淋巴细胞培养上清液总IgG 含量ELISA 检测方法的建立[J].现代生物医学进展,2012,12(26):5015-5019.
作者姓名:单纯筱王宇尹磊淼徐玉东魏颖冉君刘晓燕刘奇杨永清
作者单位:上海中医药大学上海市针灸经络研究所
基金项目:国家自然科学基金(81001548;81173341;81173332;30873286);上海市教委和上海市教育发展基金会“晨光计划”资助项目(10CG45);上海市卫生局青年基金(2009Y096);国家中医药管理局、上海市重点学科建设项目(S30304)
摘    要:目的:建立一种定量测定大鼠淋巴细胞培养上清液中总IgG(免疫球蛋白G)含量的双抗体夹心ELISA(酶联免疫吸附试验)检测方法。方法:用方阵实验确定包被抗体、检测抗体的最佳工作浓度;绘制标准曲线,确定线性范围;评价标准曲线的可重复性、精密度和可应用性。结果:包被抗体和检测抗体的最佳效价分别为2μg/ml和1:4000稀释;检测的线性范围为0.25-16ng/ml。经方法学评价,可重复性和精密度较高,应用性较强。结论:该方法灵敏度高,重复性好,可作为科研过程中检测大鼠淋巴细胞培养上清液总IgG含量的一种精确、方便、可靠的方法。

关 键 词:大鼠淋巴细胞培养上清液  IgG  双抗体夹心ELISA

Establishment of ELISA for Total IgG Content of Rat Lymphocyte Culture Supernatant
SHAN Chun-xiao,WANG Yu,YIN Lei-miao,XU Yu-dong,WEI Ying,RAN Jun,LIU Xiao-yan,LIU Qi,YANG Yong-qing.Establishment of ELISA for Total IgG Content of Rat Lymphocyte Culture Supernatant[J].Progress in Modern Biomedicine,2012,12(26):5015-5019.
Authors:SHAN Chun-xiao  WANG Yu  YIN Lei-miao  XU Yu-dong  WEI Ying  RAN Jun  LIU Xiao-yan  LIU Qi  YANG Yong-qing
Institution:△(Shanghai Research Institute of Acupuncture and Meridian,Shanghai,200030,China)
Abstract:Objective: To establish a quantitative double-antibody sandwich ELISA detecting method for total IgG content of cultured rat-lymphocyte supernatant.Methods: Detect the optimal concentrations of coated antibody and enzyme-antibody with a matrix experiment.Construct a calibration curve and determine the linear rang.Evaluation repeatability,precision and applicability of the calibration curve.Results: The optimal concentration of coated antibody is 2 μg/ml and the optimal dilution ratio of enzyme-antibody is 1:4000.The linear rang of calibration curve is 0.25-16 ng/mL.The repeatability,precision and applicability are excellent.Conclusion: This ELISA detecting method is convenient and reliable to detect total IgG content of cultured rat-lymphocyte supernatant.
Keywords:Cultured rat-lymphocyte supernatant  IgG  Double-antibody sandwich ELISA
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