A FILTER-PAPER DYE-BINDING ASSAY FOR QUANTITATIVE-DETERMINATION OF PROTEIN WITHOUT INTERFERENCE FROM REDUCING AGENTS OR DETERGENTS
A FILTER-PAPER DYE-BINDING ASSAY FOR QUANTITATIVE-DETERMINATION OF PROTEIN WITHOUT INTERFERENCE FROM REDUCING AGENTS OR DETERGENTS
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DOI：
10.1016/0003-2697(90)90134-u
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发表时间：
1990-10-01
影响因子：
2.9
通讯作者：
BAMBURG, JR
中科院分区：
文献类型：
--
作者：
MINAMIDE, LS;BAMBURG, JR
描述了一种在存在还原剂、去垢剂和其他经常干扰溶液中蛋白质测定的物质的情况下定量蛋白质的方法。将蛋白质涂在 Whatman 1 号滤纸上，风干，用甲醇洗涤，然后用考马斯亮蓝 G 染色。脱色后，将纸风干并提取蛋白质结合染料。使用自动酶标仪 (600-405 nm) 在 96 孔板中或在 610 nm 比色皿中进行样品吸光度测量。该滤纸测定可用于在存在硫酸铵、尿素、硫醇还原剂、氨基酸、DNA、离子和非离子去污剂以及酸或碱的情况下测定100ng至20μg的蛋白质。
A method is described for quantitation of protein in the presence of reducing agents, detergents, and other substances which often interfere with assays of protein in solution. The proteins are applied to Whatman No. 1 filter paper, air-dried, washed with methanol, and then stained with Coomassie brilliant blue G. Following destaining, the paper is air-dried and the protein-bound dye is extracted. Sample absorbance measurements are made in a 96-well plate using an automated microplate reader (600-405 nm) or in a cuvette at 610 nm. This filter paper assay is useful for determining 100 ng to 20 .mu.g of protein in the presence of ammonium sulfate, urea, thiol-reducing agents, amino acids, DNA, ionic and nonionic detergents, and acid or base.