ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS - PRINCIPLE, METHODOLOGY AND SOME APPLICATIONS
ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS - PRINCIPLE, METHODOLOGY AND SOME APPLICATIONS
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DOI：
10.1016/0165-022x(82)90013-6
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发表时间：
1982-01-01
影响因子：
--
通讯作者：
POSTEL, W
中科院分区：
文献类型：
--
作者：
BJELLQVIST, B;EK, K;POSTEL, W
本文介绍了一种在等电聚焦中产生pH梯度的新技术，其原理是缓冲基团共价连接到用作抗对流介质的基质上。为了在聚丙烯酰胺凝胶中产生这种类型的pH梯度，使用一组称为Immobiline（与Ampholine类似）的缓冲单体。以与孔梯度凝胶相同的方式浇铸pH梯度凝胶，但不是改变丙烯酰胺含量，而是借助于Immobiline缓冲液将轻溶液和重溶液调节至不同的pH值。可用的物种使得有可能在pH 3和10之间产生任何窄的线性pH梯度。描述了这些类型的梯度在等电聚焦[蛋白质]中的行为。与载体两性电解质产生的pH梯度相比，固定化pH梯度显示出许多优点。最重要的是：阴极漂移被完全消除;它们提供更高的分辨率和更高的负载容量;它们具有均匀的电导率和缓冲容量;并且它们代表已知和受控离子强度的环境。
A new technique for generating pH gradients in isoelectric focusing is described, based on the principle that the buffering groups are covalently linked to the matrix used as anticonvective medium. For the generation of this type of pH gradient in polyacrylamide gels, a set of buffering monomers, called Immobiline (in analogy with Ampholine), is used. The pH gradient gels are cast in the same way as pore gradient gels, but instead of varying the acrylamide content, the light and heavy solutions are adjusted to different pH values with the aid of the Immobiline buffers. Available species make it possible to generate any narrow linear pH gradient between pH 3 and 10. The behavior of these types of gradients in isoelectric focusing [of proteins] is described. Immobilized pH gradients show a number of advantages compared with carrier ampholyte generated pH gradients. The most important are: the cathodic drift is completely abolished; they give higher resolution and higher loading capacity; they have uniform conductivity and buffering capacity; and they represent a milieu of known and controlled ionic strength.