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Interference of some aqueous two-phase system phase-forming components in protein determination by the Bradford method

机译:通过Bradford方法干扰一些含水两相系统相相组分的蛋白质测定

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摘要

The interference of some specific aqueous two-phase system (ATPS) phase-forming components in bovine serum albumin (BSA) determination by the Bradford method was investigated. For this purpose, calibration curves were obtained for BSA in the presence of different concentrations of salts and polymers. A total of 19 salts [Na2SO4, (NH4)(2)SO4, MgSO4, LiSO4, Na2HPO4, sodium phosphate buffer (pH 7.0), NaH2PO4, K2HPO4, potassium phosphate buffer (pH 7.0), KH2PO4, C6H8O7, Na3C6HSO7, KCHO2, NaCHO2, NaCO3, NaHCO3, C2H4O2, sodium acetate buffer (pH 4.5), and NaC2H3O2] and 7 polymers [PEG 4000, PEG 8000, PEG 20000, UCON 3900, Ficoll 70000, PES 100000, and PVP 40000] were tested, and each calibration curve was compared with the one obtained for BSA in water. Some concentrations of salts and polymers had considerable effect in the BSA calibration curve. Carbonate salts were responsible for the highest salt interference, whereas citric and acetic acids did not produce interference even in the maximum concentration level tested (5 wt%). Among the polymers, UCON gave the highest interference, whereas Ficoll did not produce interference when used in concentrations up to 10 wt%. It was concluded that a convenient dilution of the samples prior to the protein quantification is needed to ensure no significant interference from ATPS phase-forming constituents. (C) 2011 Elsevier Inc. All rights reserved.
机译:一些特定的水性两相体系的干扰(ATPS)相形成在牛血清白蛋白(BSA)通过Bradford方法测定部件进行了研究。为了这个目的,在不同浓度的盐和聚合物的存在下获得的BSA的校正曲线。共有19克的盐[硫酸钠,(NH 4)(2)SO 4,MgSO 4干燥,LiSO4,磷酸氢二钠,磷酸钠缓冲液(pH 7.0),磷酸二氢钠,磷酸氢二钾,磷酸钾缓冲液(pH 7.0),KH 2 PO 4,C6H8O7,Na3C6HSO7,KCHO2, NaCHO2,碳酸钠对芦笋,NaHCO 3洗涤,C2H4O2,乙酸钠缓冲液(pH 4.5),以及] NaC2H3O2和7层的聚合物[PEG 4000,PEG 8000,PEG 20000,UCON 3900,聚蔗糖70000,PES 100000,和PVP 40000]进行了测试,并且每个校准曲线用在水中BSA获得的一个相比较。的盐和聚合物的一些浓度曾在BSA标准曲线相当大的影响。碳酸盐分别负责最高盐干扰,而柠檬酸和乙酸甚至没有在所测试的最大浓度水平(5重量%)产生的干扰。在聚合物中,UCON给予最高的干扰,而菲浓度达到使用10%(重量)时没有产生干扰。得出的结论是需要之前的蛋白质定量的样品的方便的稀释,以确保从ATPS相形成组分没有显著干扰。 (c)2011年elselvier Inc.保留所有权利。

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