A high-affinity reversible protein stain for Western blots
A high-affinity reversible protein stain for Western blots
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DOI：
10.1016/j.ab.2004.02.049
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发表时间：
2004-06-15
影响因子：
2.9
通讯作者：
Mallia, AK
中科院分区：
文献类型：
--
作者：
Antharavally, BS;Carter, B;Mallia, AK
我们描述了一种可逆染色技术，使用 MemCode，这是一种可逆蛋白质染色剂，通过该技术可以在硝酸纤维素和聚偏二氟乙烯 (PVDF) 膜上可视化蛋白质，而无需永久固定在膜本身上。这使得随后能够对蛋白质进行免疫印迹分析。该程序仅适用于硝酸纤维素膜和 PVDF 膜上的蛋白质印迹。 MemCode 是一种可逆蛋白质染色剂，由铜组成，作为与蛋白质非共价相互作用的有机复合物的一部分。 MemCocle 表现出快速的蛋白质染色，需要 30 秒到 1 分钟才能完成。该方法简单，应用条件方便，与基质材料和蛋白质相容。该染色比任何先前描述的基于染料的通用蛋白质染色系统更敏感。绿松石蓝色染色的蛋白质条带不会随着时间的推移而褪色，并且与丽春红 S 染色的蛋白质条带相比更容易拍照。光谱蓝色区域的吸光度为照片记录提供了良好的特性，并避免了常见生物发色团的干扰。使用 MemCode 污渍橡皮擦，硝酸纤维素膜上的蛋白质染色可在 2 分钟内轻松逆转，PVDF 膜上的染色可在 10 分钟内轻松逆转。该染色剂与一般的蛋白质印迹检测系统兼容，并且用 MemCode 染色剂进行膜处理不会干扰使用辣根过氧化物和碱性磷酸酶底物的传统化学发光或显色检测。该染色剂还适用于蛋白质的 N 末端序列分析。 (C) 2004 Elsevier Inc. 保留所有权利。
We describe a reversible staining technique, using MemCode, a reversible protein stain by which proteins can be visualized on nitrocellulose and polyvinylidine fluoride (PVDF) membranes without being permanently fixed to the membrane itself. This allows subsequent immunoblot analysis of the proteins to be performed. The procedure is applicable only to protein blots on nitrocellulose and PVDF membranes. MemCode is a reversible protein stain composed of copper as a part of an organic complex that interacts noncovalently with proteins. MemCocle shows rapid protein staining, taking 30 s to 1 min for completion. The method is simple and utilizes convenient application conditions that are compatible with the matrix materials and the protein. The stain is more sensitive than any previously described dye-based universal protein staining system. The turquoise-blue-stained protein bands do not fade with time and are easy to photograph compared to those stained with Ponceau S. Absorbance in the blue region of the spectrum offers good properties for photo documentation and avoids interference from common biological chromophores. The stain on the protein is easily reversible in 2 min for nitrocellulose membrane and in 10 min for PVDF membrane with MemCode stain eraser. The stain is compatible with general Western blot detection systems, and membrane treatment with MemCode stain does not interfere with conventional chemiluminescent or chromogenic detection using horseradish peroxide and alkaline phosphatase substrates. The stain is also compatible with N-terminal sequence analysis of proteins. (C) 2004 Elsevier Inc. All rights reserved.