自制显示系统检测免疫印迹中的目的蛋白  

Determinate of Target Protein Immunoblotting with Self-made Display Systems

作　　者：张晓楠[1] 陈南春[1] 程司堃[2] 张延凤[3] 

机构地区：[1]中国人民解放军第四军医大学生物化学与分子生物学教研室,西安710032 [2]中国人民解放军第四军医大学免疫学教研室,西安710032 [3]中国人民解放军第四军医大学药理学教研室,西安710032

出　　处：《中国生物工程杂志》2006年第2期59-61,共3页China Biotechnology

摘　　要：目前蛋白免疫印迹显示系统有显色法和化学发光法，因化学发光法具有灵敏、快速、准确等特点而倍受科研工作者的青睐，但一般商品化的化学发光试剂盒价格颇为昂贵，为此，在参考文献的基础上作了一些改进，将Tris-CI的pH值从7．5提高到11．0，同时提高了对碘苯酚的浓度（从1mmol／L提高到2mmol／L），得到了自制的化学发光试剂。首先用其检测了大肠杆茵中表达的2种不同分子量的蛋白（gam，13kDa；bet，30kDa），发现当蛋白上样量为2～20ng时，各蛋白条带均清晰显示。随后的斑点免疫印迹实验表明：自制及进口化学发光试剂在检测酶标二抗效价方面有着同样高的敏感性。At present , display systems of protein immunoblotting have two kinds of method, including show the color and chemiluminescence method. Many researchers like the chemiluminescence, because it has the fast, sensitive and accurate characteristics . However, the price of commercial reagent is very expensive. Therefore, some improvements have been made on the base of reference, enhanced Tris-C1 pH from 7. 5 to 11. 0. Simultaneously , enhanced the concentration of iodophenol from l mmol/L to 2mmol/L. So the self-made chemiluminescence reagent was obtained, first to use it for examining the different molecular weight protein which expressed in E. coli, the strip is clear when the quantity of protein is 2-20ng. Then experiment of spot immunoblotting indicated that the self-made and the import chemiluminescence reagent have the similarly high sensitivity in the examination of titer of enzyme sign the second antibody.

关 键 词：免疫印迹 辣根过氧化物酶 WESTERN BLOT gam蛋白 bet蛋白 

分 类 号：Q393.91[生物学—遗传学]