Assays for Total Protein
Assays for Total Protein
复制标题
总蛋白测定
DOI：
10.1002/9780471729259.mca03as00
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发表时间：
1995
影响因子：
--
通讯作者：
D. Matulis
中科院分区：
文献类型：
--
作者：
R. Lovrien;D. Matulis
本单元描述了三种用于定量总蛋白的铜基测定法：双缩脲法、Lowry法的变体（Hartree-Lowry法）和二喹啉酸（BCA）测定法。将蛋白质的酸水解与茚三酮检测结合以定量样品的氨基酸含量。紫外分光光度法用于测量总蛋白质并评价样品中是否存在污染物。考马斯染料结合或Bradford测定是一种非常简单的测定，并且通常非常敏感，尽管它有时会根据蛋白质在酸性pH下与染料结合的程度而给出可变的响应。最后，使用干重测量来定量纯蛋白质。支持方案描述了用于酸水解的热封玻璃管、聚丙烯酰胺凝胶威尔斯孔中的样品透析以去除低分子量污染物，以及TCA沉淀以沉淀和浓缩蛋白质并去除低分子量污染物。
This unit describes three copper‐based assays to quantitate total protein: the biuret method, a variation of the Lowry method (Hartree‐Lowry method), and the bicinchoninic acid (BCA) assay. Acid hydrolysis of a protein is coupled with ninhydrin detection to quantitate amino acid content of a sample. Ultraviolet spectrophotometry is used to measure total protein and evaluate samples for the presence of contaminants. The Coomassie dye binding, or Bradford, assay is a quite simple assay and frequently is quite sensitive, although it sometimes gives a variable response depending on how well or how poorly the protein binds the dye in acid pH. Finally, dry weight measurement is used to quantitate pure protein. Support protocols describe heat sealing glass tubes for acid hydrolysis, sample dialysis in polyacrylamide gel wells to remove low‐molecular‐weight contaminants, and TCA precipitation to precipitate and concentrate proteins and remove low‐molecular‐weight contaminants.
一种根据酶消化物的吸收光谱测定蛋白质浓度的新方法。
影响因子：
2.9
作者：
Bewley,TA
通讯作者：
Bewley,TA
影响因子：
--
作者：
G. L. Peterson
通讯作者：
G. L. Peterson