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一种具有增强银染特性的用于蛋白质电泳的机械强度高的基质。

A mechanically strong matrix for protein electrophoresis with enhanced silver staining properties.

作者信息

Patton W F, Lopez M F, Barry P, Skea W M

机构信息

Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.

出版信息

Biotechniques. 1992 Apr;12(4):580-5.

PMID:1503760
Abstract

Duracryl is a mechanically strong and elastic acrylamide-based matrix, useful for a wide variety of electrophoretic applications. The matrix is stable as a refrigerated solution for one year. Upon addition of appropriate catalysts, Duracryl forms a polymer-reinforced polyacrylamide gel matrix suitable for electrophoresis. The polymer-reinforced gel is superior to conventional polyacrylamide gels in terms of mechanical strength, elasticity and protein silver staining properties. Protein detection sensitivity by silver staining, as well as the linear response of silver deposition versus protein load, is equivalent to standard acrylamide/N,N'-methylene bisacrylamide gels. Additionally, the silver staining properties of the Duracryl matrix result in proteins appearing as monochromatic shades of grey instead of red, brown and yellow, as is the case of conventional polyacrylamide matrices. Monochromatic shades of grey are more suitable for image analysis and densitometry. The matrix is compatible with standard electroblotting and protein N-terminal sequencing procedures. Low acrylic acid content and conductivity allow incorporation of the matrix into isoelectric focusing gels. The matrix was found not to alter polypeptide migration relative to the standard acrylamide/N,N'-methylene bisacrylamide matrix.

摘要

Duracryl是一种机械强度高且具有弹性的基于丙烯酰胺的基质,适用于多种电泳应用。该基质作为冷藏溶液可稳定保存一年。加入适当的催化剂后,Duracryl会形成一种适用于电泳的聚合物增强聚丙烯酰胺凝胶基质。这种聚合物增强凝胶在机械强度、弹性和蛋白质银染特性方面优于传统聚丙烯酰胺凝胶。通过银染进行蛋白质检测的灵敏度,以及银沉积与蛋白质上样量的线性响应,与标准丙烯酰胺/N,N'-亚甲基双丙烯酰胺凝胶相当。此外,Duracryl基质的银染特性使蛋白质呈现为单色灰色阴影,而不像传统聚丙烯酰胺基质那样呈现红色、棕色和黄色。单色灰色阴影更适合图像分析和光密度测定。该基质与标准电印迹和蛋白质N端测序程序兼容。低丙烯酸含量和电导率允许将该基质掺入等电聚焦凝胶中。发现该基质相对于标准丙烯酰胺/N,N'-亚甲基双丙烯酰胺基质不会改变多肽的迁移。

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