Protein sample preparation for tissue distribution study
Protein sample preparation for tissue distribution study
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用于组织分布研究的蛋白质样品制备
DOI：
10.1002/prca.202200088
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发表时间：
2022-11-16
影响因子：
2
通讯作者：
Liu，Pingsheng
中科院分区：
文献类型：
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作者：
Cheng，Linjia;Xu，Yilu;Liu，Pingsheng
目的蛋白质在动物组织中的分布和表达水平可以指示其可能的作用。实验设计在制备过程中，为了充分溶解并保持组织中几乎所有蛋白质的稳定性，同时避免大部分影响蛋白质检测的污染物，采用2×SDS样品缓冲液（2×SDS Sample Buffer），结果本研究提供了一种相对简单、重复性好、适用范围广的蛋白质分布研究方法，其中可以克服在样品制备过程中由蛋白质降解和修饰以及由其它细胞组分如中性脂质和糖原的测定干扰引起的问题。此外，该方法通过相等的湿组织质量来表示蛋白质含量，这是表示蛋白质在各种组织中的表达水平的更好方法。可以制备几乎所有组织的高质量蛋白质样本。结论和临床相关性产生的样本适合通过蛋白质印迹法进行组织分布分析，并用于银/考马斯染色、蛋白质组学和其他蛋白质分析，这将有助于潜在的生物标志物或疾病的治疗方法。
PurposeThe distribution and expression level of a protein among animal tissues is indicative of its possible roles. It is important to establish a generally applicable method to prepare protein samples with high‐quality and achieve near 100% recovery of proteins from animal tissues.Experimental DesignDuring preparation, to sufficiently dissolve and maintain stability of almost all proteins from tissues, as well as to avoid most contaminations affecting protein detection, 2×SDS Sample Buffer, sonication and trichloroacetic acid precipitation are applied.ResultsHere we provide a relatively simple, reproducible, and broadly applicable method for studying protein distribution in most tissues, in which the issues resulting from protein degradation and modification during sample preparation and assay interference by other cellular components like neutral lipids and glycogen could be overcome. Furthermore, this method represents the protein content by equal wet tissue mass, which is a better means to present the expression level of a protein in various tissues. High‐quality protein samples from almost all tissues could be prepared.Conclusions and Clinical RelevanceThe samples produced are amenable to tissue distribution analysis by Western blotting and for silver/Coomassie staining, proteomics, and other protein analyses, which would contribute to potential biomarkers or treatments for a disease.