PDF(Pubmed)
Abstract：
In conventional cell lysate protocols, cell debris is typically discarded to obtain a cleaner lysate. However, this approach has limitations, as it may overlook vital cellular components. By discarding cell debris, researchers may inadvertently exclude crucial elements. Retaining all cellular components offers several advantages for studying molecular biology within various cellular compartments. Firstly, it provides a more accurate representation of the cellular environment. Secondly, it enables the study of complex cellular interactions, including those involving cellular structures and signaling pathways associated with debris. This shift in perspective highlights the importance of a holistic approach to lysate preparation. By obtaining lysates that include all cellular components, researchers can gain deeper insights into cellular processes, leading to more accurate data and a better understanding of cellular function and dysfunction. This study aimed to develop a protocol for the preparation of total cell lysates that retain all cellular components, including debris. Our method involves:•A three-step solubilization process using a combination of detergents, saccharides, and chelators, coupled with sonication, in contrast to the classical one-step approach using an all-detergent cocktail.•A comprehensive strategy ensuring the solubilization of all cellular components, providing a more complete lysate for analysis.
摘要：
在常规细胞裂解物方案中，细胞碎片通常被丢弃以获得更清洁的裂解物。然而,这种方法有局限性，因为它可能忽略了重要的细胞成分。通过丢弃细胞碎片，研究人员可能无意中排除了关键因素。保留所有细胞组分为研究各种细胞区室中的分子生物学提供了几个优点。首先,它提供了蜂窝环境的更准确的表示。其次,它能够研究复杂的细胞相互作用，包括与碎片相关的细胞结构和信号通路。这种观点的转变凸显了整体方法制备裂解物的重要性。通过获得包括所有细胞成分的裂解物，研究人员可以更深入地了解细胞过程，导致更准确的数据和更好地理解细胞功能和功能障碍。这项研究旨在开发一种保留所有细胞成分的总细胞裂解物的制备方案，包括碎片.我们的方法包括：•使用洗涤剂组合的三步增溶过程，糖类，和螯合剂，再加上超声处理，与使用全洗涤剂鸡尾酒的经典一步法相反。•确保所有细胞成分溶解的综合策略，提供更完整的裂解物进行分析。
