对印迹上其他显著蛋白质检测方法的简要综述。

A brief review of other notable protein detection methods on blots.

作者信息

Kurien Biji T, Scofield R Hal

机构信息

Oklahoma Medical Research Foundation, 825 NE 13th Street, OK 73104, Oklahoma City, USA.

出版信息

Methods Mol Biol. 2009;536:557-71. doi: 10.1007/978-1-59745-542-8_56.

DOI:10.1007/978-1-59745-542-8_56

PMID:19378092

Abstract

Several methods have been used for detecting proteins on membranes. These include the use of quantum dot luminescent labels, oxyblot immunochemical detection, polymer immunocomplexes, "coupled" probing approach, in situ renaturation of proteins for detecting enzyme activities in crude or purified preparations, immunochromatographic assay, western-phosphatase assay, and the use of Congo red dye, a cosmetic color named Alta, Pro-Q Emerald 488 dye, or amine-reactive dye in combination with alkaline phosphatase and horseradish peroxidase-antibody conjugates for the simultaneous trichromatic fluourescence detection of proteins. Several methods have been used to improve the detection of proteins on membranes, including glutaraldehyde treatment of nitrocellulose blots, elimination of keratin artifacts in immunoblots probed with polyclonal antibodies, and the washing of immunoblots with excessive water and manipulation of Tween-20 in wash buffer. These methods are briefly reviewed in this chapter.

摘要

已有多种方法用于检测膜上的蛋白质。这些方法包括使用量子点发光标记、氧印迹免疫化学检测、聚合物免疫复合物、“偶联”探测方法、用于检测粗提物或纯化物中酶活性的蛋白质原位复性、免疫色谱分析、western-磷酸酶分析，以及使用刚果红染料、一种名为Alta的化妆品色素、Pro-Q Emerald 488染料，或与碱性磷酸酶和辣根过氧化物酶-抗体偶联物结合使用的胺反应性染料，以同时进行蛋白质的三色荧光检测。已有多种方法用于改进膜上蛋白质的检测，包括用戊二醛处理硝酸纤维素印迹、消除用多克隆抗体探测的免疫印迹中的角蛋白假象，以及用过量水洗涤免疫印迹并在洗涤缓冲液中处理吐温-20。本章将对这些方法进行简要综述。