RAPID AND SENSITIVE METHOD FOR QUANTITATION OF MICROGRAM QUANTITIES OF PROTEIN UTILIZING PRINCIPLE OF PROTEIN-DYE BINDING
RAPID AND SENSITIVE METHOD FOR QUANTITATION OF MICROGRAM QUANTITIES OF PROTEIN UTILIZING PRINCIPLE OF PROTEIN-DYE BINDING
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DOI：
10.1016/0003-2697(76)90527-3
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发表时间：
1976-01-01
影响因子：
2.9
通讯作者：
BRADFORD, MM
中科院分区：
文献类型：
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作者：
BRADFORD, MM
本文介绍了考马斯亮蓝G-250与蛋白质结合测定蛋白质的方法。染料与蛋白质的结合使染料的最大吸收从465 nm移至595 nm，并监测595 nm处吸收的增加。该测定法具有重现性和快速性，染料结合过程几乎在约2 min内完成，4 h内颜色稳定性良好。钠或钾等阳离子以及蔗糖等碳水化合物的干扰很小或没有。在存在强碱性缓冲剂的情况下显色少量，但通过使用适当的缓冲液对照品可准确运行测定。在测定中产生过量干扰颜色的唯一组分是相对大量的洗涤剂，例如十二烷基硫酸钠、Triton X-100和商业玻璃器皿洗涤剂。少量洗涤剂的干扰可以通过使用适当的控制来消除。[使用牛血清白蛋白、马心细胞色素c和人血清白蛋白]。
A protein determination method involving the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein shifts the absorption maximum of the dye from 465 to 595 nm, and the increase in absorption at 595 nm is monitored. This assay is reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 4 h. There is little or no interference from cations such as Na or K nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components giving excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100 and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls. [Bovine serum albumin, horse heart cytochrome c and human serum albumin were used].