病毒抗原和抗体的Western印迹法及斑点免疫印迹分析：应用于小鼠肝炎病毒
Western and dot immunoblotting analysis of viral antigens and antibodies: application to murine hepatitis virus.
作者信息
Talbot P J, Knobler R L, Buchmeier M J
出版信息
J Immunol Methods. 1984 Oct 12;73(1):177-88. doi: 10.1016/0022-1759(84)90043-7.
Abstract
Viral proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred quantitatively to nitrocellulose by electroblotting in SDS-containing buffer. Monoclonal antibodies directed against previously defined epitopes on the viral proteins were used as probes to detect viral protein synthesis and processing, as well as expression in animal tissues. Circulating polyclonal antibodies were also probed and characterized for their polypeptide specificities. Under appropriate conditions, this Western immunoblotting technique was quantitative. Finally, a highly sensitive dot immunoblotting assay was used to analyze the sensitivity to denaturation of various epitopes on the viral proteins. This assay detected picogram quantities of viral antigens and antibodies.
摘要
病毒蛋白通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）进行分离，并在含SDS的缓冲液中通过电印迹法定量转移至硝酸纤维素膜上。针对病毒蛋白上先前确定的表位的单克隆抗体用作探针，以检测病毒蛋白的合成、加工以及在动物组织中的表达。还对循环多克隆抗体进行了检测，并对其多肽特异性进行了表征。在适当条件下，这种蛋白质免疫印迹技术是定量的。最后，使用一种高度灵敏的斑点免疫印迹测定法来分析病毒蛋白上各种表位对变性的敏感性。该测定法可检测皮克量的病毒抗原和抗体。
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