ELISA测定中TMB显色体系的优化及其稳定性研究-生物技术通报.PDFVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 2 ELISA TMB 李红梅 陈佳 徐斐 曹慧 M aria Eugen ia ( , 200093) : 通过对间接酶联免疫测定中显色体系的各影响因素的 究, 确定了酶联免疫测定的最优显色体系为二甲基亚 砜( DM SO)配制四甲基联苯胺( TM B)浓度为 0. 3 m g/mL 的溶液为A 液, pH 5. 5, 0. 2 m l/L磷酸氢二钠与 0. 1 m l/L柠檬酸缓 冲液配制过氧化脲素浓度为0. 74 m g/mL 的溶液为 B 液, 两者按一定比例混合后使用试验结果表明, 此显色体系在 25 - 40 内进行都可得到较为准确的结果, 且 AB 液混合后在室温下放置4 h仍可使用A 液及 B 液分别在 4 放置 60 d后混合 使用仍不影响测定结果 : 酶联免疫测定 显色 辣根过氧化物酶 TM B Optim ization of TMB Substrate Chromogenic System in ELISA and the Study of Its Stability in Storage LiH ngm ei Chen Jia Xu Fei Ca Hu i M aria Eugen ia ( S ch oo l of M ed ica l Instrum en t and F ood E ng ineering, Un iversity of Sh ang ha if or Sc ience and T echnology, Sh angha i 200093) A bstrac:t By the study f the fact rs wh ich influence the chr m gen ic reacti n in ind irect EL ISA, the ptmi al chr m gen ic system w as btained. S luti n A w as prepared by TMB diss lved in DM SO w ith the c ncentrati n f 0. 3 m g/mL, s luti n B w as prepared by hydr gen per x ide urea d iss lved in pH 5. 5, 0. 2 m l/L Na H PO - 0. 1 m l/L citric acid bufferw ith the c ncentrati n f 0. 74 m g/ 2 4 mL, and the substrate chr m gen ic system was prepared bym ix ingA and B in certain rati. The expermi ent resu lts sh w ed that this sub strate chr m gen ic system c u ld indicate the result accurately, and A, B m ixturewas still effective after 4 h urs st rage at r m tem pera tu re. Them ixture f s luti n A and B still c u ld be ab le t use in ELISA after being st red in 4 f r 60 days. K ey w ords: ELISA Chr m gen ic HRP TMB ( ELISA ) , ELISA ( HRP ) [ 1]