从单个组织样本中测量脂质介质、蛋白质和信使核糖核酸的方法。
Method for measuring lipid mediators, proteins, and messenger RNAs from a single tissue specimen.
作者信息
Cottrell Jessica A, Lin Hsuan-Ni, O'Connor J Patrick
机构信息
Department of Biochemistry and Molecular Biology, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA.
Department of Biochemistry and Molecular Biology, Graduate School of Biomedical Sciences, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA.
出版信息
Anal Biochem. 2015 Jan 15;469:34-42. doi: 10.1016/j.ab.2014.10.004. Epub 2014 Oct 20.
This article describes a new method for extracting RNA, protein, and lipid mediators from a single tissue specimen. Specifically, mouse bone fracture callus specimens were extracted into a single solution that was processed using three different procedures to measure messenger RNA (mRNA) levels by reverse transcription-quantitative polymerase chain reaction (RTqPCR), cytokines and growth factors using an xMAP method, and lipid mediators by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method has several advantages because it decreases the number of animals necessary for experimentation, allows division of the sample from a homogeneous mixture that reduces sample variability, and uses a solution that protects the integrity of the macromolecules during storage.
本文描述了一种从单个组织样本中提取RNA、蛋白质和脂质介质的新方法。具体而言，将小鼠骨折痂样本提取到单一溶液中，该溶液通过三种不同程序进行处理，以通过逆转录定量聚合酶链反应（RTqPCR）测量信使核糖核酸（mRNA）水平，使用xMAP方法测量细胞因子和生长因子，并通过液相色谱-串联质谱法（LC-MS/MS）测量脂质介质。该方法具有多个优点，因为它减少了实验所需的动物数量，允许从均匀混合物中分割样本以降低样本变异性，并且使用一种在储存期间保护大分子完整性的溶液。
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