Cell lysates is generated by lysing whole cells of a cell population using chemical agents, enzymes, or physical disruption. Cell lysates contain cell membrane, cytoplasmic and nuclear proteins, and extracts, which can be used for the purification or analysis of whole organelles, nucleic acids (genomic DNA and RNA), and cellular proteins. In addition, lysates of cell line containing either labeled or unlabeled overexpressed proteins are used as positive controls or negative controls in Western blotting and ELISA to study antibody reactivity.
There are many methods to prepare cell lysates by lysis of cells, which are mainly divided into mechanical methods and non-mechanical methods. In mechanical methods, the cell membrane is physically broken down by shear or external force. Mechanical methods are controllable and direct cellular disruption methods, but some mechanical methods are not easy to operate. Non-mechanical methods are gentler than mechanical methods, but some components of the lysis solution may affect downstream experiments and needs to be removed prior to use. In non-mechanical methods, a specially formulated lysis buffer containing certain chemicals or enzymes is used to disrupt cell membranes. Enzymes commonly used for cell lysis include cellulase, protease, lysozyme, zymolyase, and glycanase. In detergent-based cell lysis buffers, surfactants cause the cell lysis by modifying the lipid organization and the integral protein arrangement. In addition, chaotropes are also used in lysis buffers, such as sodium iodide, guanidine HCl, guanidine isothiocyanate, and urea.
Cell lysates as an excellent tool and are more convenient to use than a live cell line without the need for cell culture. Cell lysates can be prepared based on specific properties such as subcellular compartments, cell lines, cell types, overexpressed proteins, or gene knock-out. Amerigo Scientific offers high-quality cell lysate products generated from a variety of cell lines.
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