立显蛋白染色液在SDS-PAGE中检测微量蛋白的应用  

Application of fluorescent staining buffer in test for trace protein in SDS-PAGE

作　　者：王世东 赵墩[1] 胡玉立[1] 周小飞 余兴龙[1] WANG Shi-dong;ZHAO Dun;HU Yu-li;ZHOU Xiao-fei;YU Xing-long(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,Hunan Province,China)

机构地区：[1]湖南农业大学动物医学院

出　　处：《中国生物制品学杂志》2019年第7期803-805,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目(31672539)

摘　　要：目的探讨立显蛋白染色液在SDS-PAGE中检测微量蛋白的应用效果。方法制备SDS-PAGE胶,采用立显蛋白染色液及普通考马斯亮蓝(Coomassie brilliant blue,CBB)染色液,检测定量的牛血清白蛋白(bovine serum albumin,BSA)的敏感度及猪细小病毒VP2蛋白、猪蓝耳病病毒N蛋白、猪丹毒杆菌Fe蛋白和猪伪狂犬病病毒gB蛋白基因工程纯化样品的纯度。结果采用立显蛋白染色液染色可识别蛋白质含量为2.5ng的蛋白条带,敏感性高于普通CBB染色40倍。4种蛋白纯化样品经CBB染色后检测纯度均较高,但经立显蛋白染色液染色后,4种纯化蛋白样品均检测出杂蛋白,测定的纯度均有所下降,其中VP2及gB蛋白的纯度有较大幅度下降(分别由83.33%、100%降至51.8%、85.6%)。结论立显蛋白染色液染色SDS-PAGE胶中的蛋白条带敏感性高,快速简便,适用于微量蛋白质的检测。Objective To investigate the application of fluorescent staining buffer in test for trace protein in SDS-PAGE.Methods SDS-PAGE gel was prepared,with which the sensitivities of fluorescent staining buffer and Coomassie brilliant blue(CBB)in determination of quantitative bovine serum albumin(BSA)were compared. On the basis of this,the purities of porcine parvovirus VP2 protein,porcine reproductive and respiratory syndrome virus N protein,Erysipelas erysipelas Fe protein and porcine pseudorabies virus gB protein determined with fluorescent staining buffer and CBB were compared.Results The protein band at a content of 2. 5 ng was detected with fluorescent staining buffer,indicating a sensitivity 40-fold higher than that with CBB. The purities of four proteins determined with CBB were relatively high. However,foreign proteins were detected in all the proteins with fluorescent staining buffer,while the purities were relatively low. Compared with that by CBB,the purities of porcine parvovirus VP2 protein and porcine pseudorabies virus gB protein determined by fluorescent staining buffer decreased from 83. 33% to 51. 8% and from 100% to 85. 6% respectively. Conclusion The fluorescent staining buffer showed high sensitivity in SDS-PAGE,which was rapid and simple,and was suitable for determination of trace protein.

关 键 词：立显蛋白染色液 考马斯亮蓝染色液 SDS-PAGE 微量蛋白 

分 类 号：R373.23[医药卫生—病原生物学] Q75[医药卫生—基础医学]