Improved semiquantitative Western blot technique with increased quantification range
Improved semiquantitative Western blot technique with increased quantification range
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DOI：
10.1016/j.jim.2009.03.018
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发表时间：
2009-06-30
影响因子：
2.2
通讯作者：
Bader, A.
中科院分区：
文献类型：
--
作者：
Heidebrecht, F.;Heidebrecht, A.;Bader, A.
随着系统生物学等新的交叉学科领域的发展，对生物样品中蛋白质表达的定量分析越来越受到重视。虽然最常见的方法是ELISA法，但Western印迹也有优点：根据大小分离蛋白质只允许评估特定结合的蛋白质。本工作研究了Western印迹信号链，确定了与定量分析相关的一些参数，并提出了反应动力学的数学模型。在此模型的基础上，建立了一种利用双曲线校正曲线同时定量不同蛋白质的半定量蛋白质印迹方法。编制了用于双曲线回归的程序，允许快速确定校准曲线系数。该程序还可用于其他应用中的校准曲线的逼近，如EL ISA、BCA或Bradford分析。(C)2009爱思唯尔B.V.保留所有权利。
With the development of new interdisciplinary fields such as systems biology, the quantitative analysis of protein expression in biological samples gains more and more importance. Although the most common method for this is ELISA, Western blot also has advantages: The separation of proteins by size allows the evaluation of only specifically bound protein. This work examines the Western blot signal chain, determines some of the parameters relevant for quantitative analysis and proposes a mathematical model of the reaction kinetics. Using this model, a semiquantitative Western blot method for simultaneous quantification of different proteins using a hyperbolic calibration curve was developed. A program was written for the purpose of hyperbolic regression that allows quick determination of the calibration curve coefficients. This program can be used also for approximation of calibration curves in other applications such as ELISA, BCA or Bradford assays. (C) 2009 Elsevier B.V. All rights reserved.