G-Bioscience Sample Preparation – Lysis – Fractionation – Clean-Up & Concentration – Handbook & Selection Guide

Gbioscience

Cell or tissue lysis, fractionation and sample preparation are crucial tools for the purification, analysis and identification of proteins and their functions or roles. Unfortunately, there is no single procedure or protocol for optimal protein sample preparation as the techniques used are dependednt on numerous factors, including starting sample and downstream analysis techniques. There are generally three main stages:1. Cell/Tissue Lysis: The release of proteins.2. Protein Fractionation: The simplification of the protein complexity by fractionation. 3. Sample Preparation: The specific clean-up, concentration and additional treatments for subsequent analysis techniques (i.e. 1D or 2D protein electrophoresis). Cell/ Tissue Lysis is the first step that is involved in cell extraction and protein purification. G-Biosciences offers a wide selection of protein extraction and lysis buffer systems. The range includes products that maintain biological activity of proteins, strong chaotropic extraction buffers that are 2D compatible and extraction systems for total proteomes. Upon release of the proteins from the cell or tissue, simplification of the protein complex is performed. Protein analysis is often inhibited by the vast amount of proteins present and the large abundant proteins often inhibit the analysis of the low abundant proteins. Researchers overcome this problem by using fractionation, however inconsistencies in techniques and buffers often results in a lack of reproducibility. To aid in the simplification of samples, G-Biosciences offers several products for the rapid fractionation of proteins into multiple characteristics, including cellular location, hydrophobicity, post-translational modifications and other protein properties. After lysis of the cell and protein fractionation has occurred, the final stage of identification of the protein, their roles and functions is to clean-up the sample for subsequent analysis techniques. G-Biosciences offers unique dialysis systems for the rapid removal of interfering agents from samples, ensuring no sample loss. Specialized clean-up kits are offered for protein samples destined for analysis by 1D and 2D electrophoresis. Several protein concentration kits are offered for the rapid concentration of dilute protein samples as well.A wide range of lysis buffers and systems are available that offer researchers a large choice of lysis conditions, including total denaturing lysis, chaotropic extraction, gentle lysis for biologically active proteins, isolation of total proteomes and more. Cell/Tissue lysis————————————————————————————A wide selection of protein extraction and lysis buffer systems are offered. The range includes products that maintain biological activity of proteins (PE LB™ systems), strong chaotropic extraction buffers that are 2D compatible (2D-Xtract™, FOCUS™ Extraction Buffers) and extraction systems for total proteomes (FOCUS™ Proteome kits).Common lysis buffers (RIPA), extraction tools (grinding resins), enzymes (lysozyme and Zymolyase®), protease and phosphatase inhibitors and other extraction accessories are also offered.ProTein exTraCTion & lysis Buffer (Pe lB™) sysTems————————————————————————————Lysis and extraction of biologically active proteins from cellular and tissue samples is the first critical step for biochemical analysis. The correct selection of lysis and extraction buffers requires knowledge of the proteins of interest and the stability of their biological activities. The Protein Extraction & Lysis Buffer (PE LB™) systems ensure good protein recovery, while maintaining the biological activity of the proteins. The solubilized proteins are suitable for enzyme assays, electrophoresis, folding studies, chromatographic studies and many other downstream applications. PE LBFigure 1: ™ System maintains the biological activity of proteins. Extraction of carbonic anhydrase or alkaline phosphatase from E.coli, human cells, yeast and mouse pancreas with Bacterial, Mammalian Cell, Yeast and Tissue PE LB™ respectively. The resulting lysates were submitted to enzyme assays and both enzymes retain their biological activity.The PE LB™ systems are based on a proprietary combination of organic buffering agents, mild non-ionic detergents, and a combination of various salts to enhance extraction of proteins and maintain stability of biological activities of the proteins. Depending on application, additional agents such as chelating agents, reducing agents and protease and phosphatase inhibitors may be added to the PE LB™ buffer system.The PE LB™ systems are compatible with most downstream applications including enzyme assays, running various chromatographic applications, gel electrophoresis applications, and protein folding procedures.

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