Gel Filtration Chromatography . Molecules move through a bed of porous beads, diffusing into the beads to greater or lesser degrees. Smaller molecules diffuse further into the pores of the beads and therefore move through the bed more slowly, while larger molecules enter less or not at all and thus move through the bed more quickly. Both molecular weight and three- dimensional shape contribute to the degree of retention. Gel Filtration Chromatography may be used for analysis of molecular size, for separations of components in a mixture, or for salt removal or buffer exchange from a preparation of macromolecules. Molecular Weight Ranges for Gel Filtration Media. Hydrophobic / Lipophilic. Hydroxypropyl beaded dextran (Lipohilic Sephadex. The HR grades are smaller particle sizes with narrower size distributions optimized for more efficient separations with faster flow. Gives fast and easy high- resolution results in both lab and industrial scales. Medium. Fraction Ranges (proteins)Properties / Limitations. Potential Applications. Matrix: acrylic (dextran/bisacrylamide copolymer)S- 1. HR: 1. K – 1. 00. KClean with 0. 2. M Na. OH, 0. 1. M HCl, 1. M HOAc. Sterilize by autoclave. S- 1. 00 HR: peptides, small proteins. Widely used in industrial processes. Also useful in desalting and buffer exchange. Medium. Fraction Ranges (proteins)Properties / Limitations. Potential Applications. Matrix: dextran (crosslinked with epichlorohydrin)G- 1. Clean with 0. 2. M Na. OHSterilize by autoclave. Principles of Gel Filtration Chromatography. Introduce chromatographic separation to your class and show them how dyes of different colors separate on the basis of their size and shape. This experiment contains. Gel filtration chromatography (sometimes referred to as size exclusion chromatography) separates biomolecules based on differences in their molecular size. The process employs a gel media suspended in. G- 1. 0: very low MW substances; desalting; buffer exchange; peptides. C: 1. 00–3. 00 . Cross- linked beaded agarose is more resistant to denaturing conditions, and thus offers more versatility in the choice of sample buffer and eluent. The approximate % agarose concentration is indicated by the first number of the Stock Number. Often used as a base matrix for coupling affinity ligands or separation of very large molecules and virus particles. Medium. Fraction Ranges (proteins)Properties / Limitations. Potential Applications. Matrix: agarose. 6B: 1. K – 4. 00. 0KClean with 0. M Na. OHCannot be autoclaved. Temperature range: 4–4. High- resolution with short run times and good recovery. Medium. Fraction Ranges (proteins)Properties / Limitations. Introduction to TSK-GEL Size Exclusion Chromatography Columns Tosoh Bioscience provides TSK-GEL columns for both modes of Size Exclusion Chromatography (SEC), which are Gel Filtration Chromatography (GFC) and Gel Permeation. 18-1022-18 Edition AI Gel Filtration Handbook – Principles and Methods www.chromatography.amershambiosciences.com Production: RAK Design AB Principles and Methods Gel Filtration. Official Full-Text Publication: Gel-Filtration Chromatography on ResearchGate, the professional network for scientists. Potential Applications. Matrix: dextran (covalently bonded to agarose)3. KClean with 1. M Na. OH, 0. 1. M HCl, 1. M HOAc. Sterilize by autoclave. Selectivity between Sephadex G- 2. G- 5. 02. 4 – 4. 4 . Selectivity similar to Sephadex. G- 7. 50. 5 MPa. 20. K – 6. 00. KCompatible with organic solvents. MW; good when protein MW is unknown. Selectivity similar to Sephadex. G- 2. 00p. H range: 3–1. Scaleable. High recoveries. Highest selectivity Visit our listing of Superdex Matrices. Superdex is a registered trademark of GE Healthcare. Superose. Highly cross- linked beaded agarose. Medium. Fraction Ranges (proteins)Properties / Limitations. Potential Applications. Matrix: agarose (highly crosslinked)1. K – 3. 00. KClean with 0. M Na. OH, 0. 0. 1MHCl, 1. M HOAc. Sterilize by autoclave. Preparative work: proteins; DNA fragments; polysaccharides. Toyopearl HW gels are synthesized from hydrophilic vinyl polymer containing numberous hydroxyl groups and are composed exclusively of C, H and O atoms. Toyopearl HW is very strong mechanically and can be used at high flow rates. Medium. Fraction Ranges (proteins)Properties / Limitations. Potential Applications. Matrix: methacrylate (ethylene glycol/methacrylate copolymer)4. KClean with surfactant, dissociating agent, acid/base. Sterilize by autoclave. HW- 4. 0: very effective with organics; phthalates; cyclodextrins; low MW nucleosides/nucleotides; peptide digests.
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