Zhang Li, Wei Qingyi, Mao Li, Liu Wenbin, Mills Gordon B, Coombes Kevin
Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, 1400 Pressler street, Unit 1410, Houston, TX 77030, USA.
Bioinformatics. 2009 Mar 1;25(5):650-4. doi: 10.1093/bioinformatics/btn663. Epub 2009 Jan 28.
Reverse phase protein arrays (RPPAs) are a powerful high-throughput tool for measuring protein concentrations in a large number of samples. In RPPA technology, the original samples are often diluted successively multiple times, forming dilution series to extend the dynamic range of the measurements and to increase confidence in quantitation. An RPPA experiment is equivalent to running multiple ELISA assays concurrently except that there is usually no known protein concentration from which one can construct a standard response curve. Here, we describe a new method called 'serial dilution curve for RPPA data analysis'. Compared with the existing methods, the new method has the advantage of using fewer parameters and offering a simple way of visualizing the raw data. We showed how the method can be used to examine data quality and to obtain robust quantification of protein concentrations.
A computer program in R for using serial dilution curve for RPPA data analysis is freely available at http://odin.mdacc.tmc.edu/~zhangli/RPPA.
反相蛋白质阵列(RPPA)是一种强大的高通量工具,用于测量大量样本中的蛋白质浓度。在RPPA技术中,原始样本通常会连续多次稀释,形成稀释系列,以扩展测量的动态范围并提高定量的可信度。RPPA实验相当于同时进行多个ELISA检测,只是通常没有已知的蛋白质浓度来构建标准响应曲线。在此,我们描述了一种名为“用于RPPA数据分析的系列稀释曲线”的新方法。与现有方法相比,新方法具有使用参数较少且提供直观呈现原始数据的简单方式的优点。我们展示了该方法如何用于检查数据质量以及获得可靠的蛋白质浓度定量。
用于RPPA数据分析的系列稀释曲线的R计算机程序可在http://odin.mdacc.tmc.edu/~zhangli/RPPA免费获取。