蛋白质芯片制作条件的优化

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (11): 1230-1232.

蛋白质芯片制作条件的优化

江凌晓¹, 郭兆彪², 陈泽良², 王津², 王红霞², 俞守义¹, 杨瑞馥²

1. 南方医科大学流行病教研室, 广东, 广州, 510515;
2. 军事医学科学院微生物流行病研究所, 北京, 100071

出版日期:2004-11-20 发布日期:2004-11-20
基金资助:
收稿日期:2004-2-18。
基金项目:国家863计划课题(2001AA223061)
作者简介:江凌晓(1972- ),女,博士,于军事医学科学院微生物流行病研究所完成课题研究,研究方向:微生物蛋白质芯片.通讯作者:杨瑞馥,电话:010-66948595,E-mail:yangrf@nic.bmi.ac.cn,俞守义,电话:020-61648312
通讯作者:杨瑞馥,电话:010-66948595,E-mail:yangrf@nic.bmi.ac.cn;俞守义,电话:020-61648312

Optimization of the conditions for protein chip preparation

JIANG Ling-xiao¹, GUO Zhao-biao², CHEN Ze-liang², WANG Jin², WAZNG Hong-xia², YU Shou-yi¹, YANG Rui-fu²

1. 南方医科大学流行病教研室, 广东, 广州, 510515;
2. 军事医学科学院微生物流行病研究所, 北京, 100071

Online:2004-11-20 Published:2004-11-20

摘要/Abstract

摘要： 目的探讨以醛基化玻片为固相载体制备蛋白质芯片的条件。方法蛋白质用含40%甘油的PBS稀释后点加在醛基化玻片上,室温干燥1 h,4℃冰箱保存,经封闭剂封闭,PBS漂洗后,依次加入抗体,反应结果用扫描仪进行扫读。结果以醛基化玻片为固相载体时预点10~15个点后点样均一性较好;点样后4℃冰箱保存24~48 h再进行漂洗和封闭处理,所得结果的荧光测量值较高;3% BSA为封闭剂时所得图像的荧光背景值最低;点样量影响荧光信号的强度。结论以醛基化玻片为固相载体制备蛋白质芯片时要预点15~20个点后再正式点样,点样后应4℃保存24~48 h再进行漂洗和3% BSA封闭处理。

Abstract: Objective To define the optimal conditions for preparing protein chips on aldehyde-coated slides.Methods The proteins were diluted in PBS containing 40% glycerol and spotted on aldehyde-coated slides. After the spots were dried for 1 hour at room temperature, the slides were stored at 4℃. Following block and rinse, the slides were used for immunoassay and the results detected with a scanner. Several key factors that might influence the results were tested, including the number of spots, concentration of protein in the spotting solution, time of immobilization and the blocking reagent.Results Pre-spotting of 10 to 15 spots achieved good homogeneity of the subsequent spots on aldehyde-coated slides. The protein immobilized at 4℃ for 24 to 48 h showed higher fluorescence intensity and clearer images, and the slides blocked with 3% BSA produced the lowest background signal. The concentration of protein in the spotting solution significantly affected the fluorescence intensity.Conclusion To ensure good results in preparing protein chips on aldehyde slides, pre-spotting of 10-15 spots can be necessary followed by immobilization at 4℃ for 24-48h and 3% BSA blocking.

中图分类号:

江凌晓¹, 郭兆彪², 陈泽良², 王津², 王红霞², 俞守义¹, 杨瑞馥². 蛋白质芯片制作条件的优化[J]. 南方医科大学学报, 2004, 24(11): 1230-1232.

JIANG Ling-xiao¹, GUO Zhao-biao², CHEN Ze-liang², WANG Jin², WAZNG Hong-xia², YU Shou-yi¹, YANG Rui-fu². Optimization of the conditions for protein chip preparation[J]. Journal of Southern Medical University, 2004, 24(11): 1230-1232.

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