OPTIMAL STAINING CONDITIONS FOR THE QUANTITATIVE ANALYSIS OF HUMAN SERUM PROTEIN FRACTIONS BY CELLULOSE ACETATE ELECTROPHORESIS
OPTIMAL STAINING CONDITIONS FOR THE QUANTITATIVE ANALYSIS OF HUMAN SERUM PROTEIN FRACTIONS BY CELLULOSE ACETATE ELECTROPHORESIS
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DOI：
10.1021/ac60166a602
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发表时间：
1960-01-01
影响因子：
7.4
通讯作者：
BRACKENRIDGE, CJ
中科院分区：
文献类型：
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作者：
BRACKENRIDGE, CJ
作为5种主要人血清蛋白组分定量电泳分析的基础，本文确定了丽丝胺绿在醋酸纤维素上染色的最佳条件。研究了pH值对染料洗脱的影响，以及pH值、离子强度、染色时间和染色温度对染料摄取的影响，并应用于测量五种血浆蛋白、一种黏液蛋白和两种脂蛋白的染料结合能力。最佳染料摄取发生在与丽丝胺绿表观解离常数相关的pH值。对染色动力学的计算表明，选择的最佳染色时间明显长于反应的半衰期。由于白蛋白的温度系数相对较高，因此染色应进行恒温控制。白蛋白和四种球蛋白的染料摄取几乎与浓度成线性关系，证实了丽丝胺绿对蛋白质染色的适用性。
As a basis for the quantitative electrophoretic analysis of five major human serum protein fractions, the optimal conditions for staining with Lissamine Green on cellulose acetate have been elucidated. The effects of pH on dye elution, and of pH, ionic strength, staining time, and staining temperature on dye uptake are ex-amined and applied to the measure-ment of dye-bindingcapacities of five plasma proteins, one mucoprotein, and two lipoprotein preparations. Optimal dye uptake occurs at a pH related to the apparent dissociation constant of Lissamine Green. Calcula-tions on the kinetics of staining indicate that the optimal staining time chosen is appreciably longer than the half life of the reaction. Staining should be thermostatically controlled because of the relatively high temperature co-efficient of albumin. The dye uptakes of albumin and four globulin fractions were virtually linearwith respect to concentration, and confirm the suit-ability of Lissamine Green for staining proteins.