Bradford 法测定植物蛋白：1. 评价与标准方法。
Measuring plant protein with the Bradford assay : 1. Evaluation and standard method.
机构信息
Institute of Ecosystem Studies, The New York Botanical Garden, Mary Flagler Cary Arboretum, Box AB, 12545, Millbrook, New York.
出版信息
J Chem Ecol. 1989 Mar;15(3):979-92. doi: 10.1007/BF01015193.
The suitability of the Bradford protein assay for measuring plant protein was evaluated and a standard method developed. The assay involves extraction of dried, fresh, or frozen plant material in 0.1 NaOH for 30 min. Replicate 100-μl aliquots of centrifuged supernatant are assayed with 5 ml Bio-Rad Bradford dye reagent (Coomassie brilliant blue G-250) diluted 1:4 and containing 3 mg/ml soluble polyvinylpyrollidone. Absorbance at 595 nm is recorded after 15 min against an NaOH blank. Samples are calibrated against a ribulose 1,5-diphosphate carboxylase-oxygenase standard in NaOH. Procedures for plant preparation, extraction stability, the effects of phenol removal and quinone formation, and assay recovery are evaluated. Assay absorbance stability and techniques for increasing absorbance stability are reported. Changes in protein quality are briefly discussed.
评价了 Bradford 蛋白测定法用于测定植物蛋白的适用性，并制定了标准方法。该测定法包括将干燥、新鲜或冷冻的植物材料在 0.1 NaOH 中提取 30 分钟。离心上清液的 100-μl 等分试样与 5 ml Bio-Rad Bradford 染料试剂（考马斯亮蓝 G-250）在 1:4 稀释，含有 3 mg/ml 可溶性聚乙烯吡咯烷酮一起进行测定。15 分钟后，在 NaOH 空白对照下记录 595nm 处的吸光度。样品用 NaOH 中的核酮糖 1,5-二磷酸羧化酶-加氧酶标准品进行校准。评估了植物制备、提取稳定性、酚去除和醌形成的影响以及测定回收率的程序。报告了测定吸光度稳定性和增加吸光度稳定性的技术。简要讨论了蛋白质质量的变化。
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