A single step and rapid protein extraction protocol developed for cell lines and tissues: Compatible for gel based and gel free proteomic approaches

溶解 蛋白质组学 裂解缓冲液 蛋白质纯化 计算生物学 萃取(化学) 样品制备 色谱法 协议(科学) 化学 生物 生物化学 医学 基因 病理 替代医学
作者
Khushman Taunk,Debasish Paul,Raju Dabhi,Chanukuppa Venkatesh,Saikiran Jajula,Venkateshwarlu Naik,Anup Sunil Tamhankar,Tufan Naiya,Manas Kumar Santra,Srikanth Rapole
出处
期刊:Methods [Elsevier BV]
卷期号:220: 29-37 被引量:5
标识
DOI:10.1016/j.ymeth.2023.10.011
摘要

Proteins are crucial research molecules in modern biology. Almost every biological research area needs protein-based assays to answer the research questions. The study of the total protein content of a biological sample known as Proteomics, is one of the highly rated qualitative and quantitative approach to address numerous biological problems including clinical research. The key step to successfully generate high quality proteomics data is the efficient extraction of proteins from biological samples. Although different methods are in use for protein extraction from a wide variety of samples, however, because of their prolonged protocol and multiple steps involved, final protein yield is sacrificed. Here, we have shown the development of a simple single step method for extraction of proteins from mammalian cell lines as well as tissue samples in an effective and reproducible manner. This method is based on lysis of samples directly in a modified lysis buffer without CHAPS (7 M Urea, 2 M Thiourea, and 10 mM Tris-Cl; pH 8.5) that is compatible with gel based and gel free approaches. This developed protocol is reliable and should be useful for a wide range of proteomic studies involving various biological samples.
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