超敏化学发光液稀释法降低蛋白印迹的发光强度Reduced Light Intensity of Western Blot by Diluted Ultra-enhanced Chemiluminescent Solution
范才文;唐娟;罗琴;李璐;向秋;
摘要(Abstract):
蛋白印迹(western blot)分析是根据抗原、抗体特异性结合的原理检测复杂样品中某蛋白的技术。然而当样品中的蛋白浓度过低,特别是目标蛋白为低丰度蛋白时,采用普通化学发光检测,有时出现蛋白条带发光弱,曝光不够,而采用超敏化学发光检测则出现蛋白条带发光强,曝光过度,两者的蛋白条带显影图像均不理想。本文将超敏化学发光液稀释后再检测,显影效果显著提高,蛋白条带图像清晰,可见超敏化学发光液稀释法降低蛋白印迹的发光强度,提高了蛋白条带的显影效果。
关键词(KeyWords): 蛋白印迹;超敏发光液;化学发光;稀释
基金项目(Foundation): 国家自然科学基金(81260344);; 广西自然科学基金(2013GXNSFAA019228);; 广西壮族自治区教育厅资助项目(LX2014274)
作者(Authors): 范才文;唐娟;罗琴;李璐;向秋;
DOI: 10.16088/j.issn.1001-6600.2017.02.016
参考文献(References):
- [1]牟宏宇,罗波,何涛.Western blotting中低丰度蛋白转膜方法改进[J].泸州医学院学报,2014,37(5):472-475.
- [2]张莉萍,蒋纪恺,JOE T.Western-blot检测蛋白表达的方法改进及初步应用[J].第三军医大学学报,2001,23(10):1242-1243.
- [3]ZANGHERI M,DI N F,MIRASOLI M,et al.Chemiluminescence lateral flow immunoassay cartridge with integrated amorphous silicon photosensors array for human serum albumin detection in urine samples[J].Analytical and Bioanalytical Chemistry,2016,408(30):8869-8879.
- [4]高重天,孙群.如何正确估计蛋白质SDS聚丙烯酰胺凝胶电泳的样品加样量[J].生物学通报,2009,44(3):54.
- [5]D’SOUZA A,SCOFIELD R H.Protein stains to detect antigen on membranes[J].Methods in Molecular Bioloby,2009,536:433-440.
- [6]LIU J,ZHANG L,FU C,et al.Employment of 4-(1,2,4-triazol-1-yl)phenol as a signal enhancer of the chemiluminescent luminol-H2O2-horseradish peroxidase reaction for detection of hepatitis C virus in real samples[J].Luminescence,2015,30(8):1297-302.
- [7]张纪磊.化学发光共振能量转移体系的研究及其在肿瘤标志物检测中的应用[D].青岛:青岛科技大学,2011.