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  • thin layer chromato
  • Measurement of the enzyme activity was done by thin layer chromatography of a radioactive substrate sensitive to the NAPE-PLD enzymatic activity: Cleavage of the substrate affected where it appeared on the plate when the radiation was detected on a bioimaging analyzer. (wikipedia.org)
  • The eluent is optimized in small scale pretests, often using thin layer chromatography (TLC) with the same stationary phase. (wikipedia.org)
  • packed gel filtration
  • When a sample solution is passed through a column of packed gel filtration resin, small molecules in the sample (buffer salts, small molecules, etc.) enter the pores of resin beads that they encounter and are forced to follow a circuitous path before later exiting the beads. (thermofisher.com)
  • elution
  • Structural changes in galactomannan on germination of lucerne, carob, honey locust, guar and soybean seeds, as measured by viscosity, elution volumes on gel filtration and ultra-centrifugation were slight consistent with a rapid and complete hydrolysis of a molecule once hydrolysis of the mannan chain starts. (megazyme.com)
  • In the most common FPLC strategy, ion exchange, a resin is chosen that the protein of interest will bind to the resin by a charge interaction while in buffer A (the running buffer) but become dissociated and return to solution in buffer B (the elution buffer). (wikipedia.org)
  • Because of the abundant separating columns, elution systems, and detectors available, chromatography has developed into the main method for ion analysis. (wikipedia.org)
  • silica gel
  • After adsorption of nonpolar substances by addition of silica gel 60 (5 g/liter, EM Science, Gibbstown, NJ), the conditioned medium was filtered, concentrated 15-fold, and exchanged into 10 mM Na 2 HPO 4 /NaH 2 PO 4 , 100 mM NaCl, 0.1 mM imidazole ( Sigma Chemical Co. ), pH 8.3. (rupress.org)
  • For example, one of the most widely used silica gel grades in the former technique is mesh 230 - 400 (40 - 63 µm), while the latter technique typically requires mesh 70 - 230 (63 - 200 µm) silica gel. (wikipedia.org)
  • chromatographic
  • Sealed chromatography cartridges or columns work similarly except the sample and buffer is pumped into and through the resin by an external device such as a liquid chromatographic (LC) system, also requiring collection and monitoring of several fractions. (wikipedia.org)
  • These highly specialized gel filtration and chromatographic media are composed of macroscopic beads synthetically derived from the polysaccharide dextran. (wikipedia.org)
  • resin
  • The method is commonly referred to as desalting when the goal is to remove buffer salts from a sample in exchange for water (with water used to pre-equilibrate the gel-filtration resin). (thermofisher.com)
  • The resin particles have a range of pore sizes that determine the size of molecules that can be separated. (thermofisher.com)
  • Thus, the difference in the flow rates of small molecules and excluded molecules allows the faster-flowing macromolecules to become separated from the slower small molecules as the sample travels the distance of the resin bed packed in the column. (thermofisher.com)
  • This is done by passing samples through a column whose resin-bed is sufficiently tall and voluminous to fully separate the emergence from the end of the column of macromolecules compared to the small molecules. (thermofisher.com)
  • Desalting and buffer exchange both entail recovering the components of a sample in whatever buffer is used to pre-equilibrate the small, porous polymer beads (resin). (wikipedia.org)
  • Desalting occurs when buffer salts and other small molecules are removed from a sample in exchange for water (with the resin being pre-equilibrated in water). (wikipedia.org)
  • However, for buffer exchange and desalting applications, it is mainly the maximum effective pore size (exclusion limit or molecular weight cut off (MWCO) of the resin) that determines the size of molecules that can be separated. (wikipedia.org)
  • By passing samples through a column resin bed with sufficient length and volume, macromolecules can be fully separated from small molecules that travel a greater distance though the pores of the resin bed. (wikipedia.org)
  • For typical desalting and buffer exchange applications choosing a resin with a molecular weight cut off between 5 and 10KDa is usually best. (wikipedia.org)
  • The macromolecular components are recovered in the buffer used to pre-equilibrate the gel-filtration matrix, while the small molecules can be collected in a later fraction volume or be left trapped in the resin. (wikipedia.org)
  • To ensure that the resin is protonated and positively charge, the chromatography should be performed at least 2 pH units below the pKa of the amine group, 10. (wikipedia.org)
  • HPLC
  • In the case of HPLC the mobile phase consists of a non-polar solvent(s) such as hexane in normal phase or a polar solvent such as methanol in reverse phase chromatography and the sample being separated. (wikipedia.org)
  • HPLC has been used for manufacturing (e.g. during the production process of pharmaceutical and biological products), legal (e.g. detecting performance enhancement drugs in urine), research (e.g. separating the components of a complex biological sample, or of similar synthetic chemicals from each other), and medical (e.g. detecting vitamin D levels in blood serum) purposes. (wikipedia.org)
  • Some HPLC techniques use water-free mobile phases (see Normal-phase chromatography below). (wikipedia.org)
  • funnel
  • Usually, the mixture is dissolved in a suitable solvent such as dichloromethane or diethyl ether (ether), and poured into a separating funnel. (wikipedia.org)
  • dialysis
  • Gel filtration chromatography is useful for many of the same purposes as dialysis, because both methods are based on similar ranges of molecular weight cut-off (MWCO) limits that exclude molecules based on size. (thermofisher.com)
  • Compared to dialysis, gel filtration has the advantage of speed (a few minutes vs. hours for dialysis), which is necessary in certain experimental situations. (thermofisher.com)
  • Also, gel filtration is compatible with organic and other solvents that dissolve or otherwise compromise the integrity of dialysis membranes. (thermofisher.com)
  • Dialysis is useful for many of the same desalting and buffer exchange applications performed with gel filtration chromatography, as both methods are based on similar molecular weight cut-off limits. (wikipedia.org)
  • Gel filtration has the advantage of speed (a few minutes vs. hours for dialysis), though, along with the ability to remove contaminants from relatively small-volume samples compared to dialysis, an important feature when working with toxic or radioactive substances. (wikipedia.org)
  • adsorbent
  • The components of the sample mixture are separated from each other due to their different degrees of interaction with the adsorbent particles. (wikipedia.org)
  • substrate
  • After chemical synthesis and liquid chromatography-tandem mass spectrometry analysis, 4-hydroxy-5-methyl-2-methylene-3(2H)-furanone was confirmed as a substrate of FaQR and the natural precursor of HDMF. (plantcell.org)
  • You may check the assay buffer components of earlier report for this enzyme, and use the buffer (without substrate) to run gel filtration. (biotechniques.com)
  • protein
  • Thank you for your attention, but my problem is that the protein supposed to be in dimer form while it is coming out of gel filtration chromatography as a monomer. (biotechniques.com)
  • If the pH of the buffer is below the pI of the protein being run, the protein will migrate to the negative pole of the gel (positive charge is attracted to the negative pole). (wikipedia.org)
  • phases
  • The stationary phases are usually finely ground powders or gels and/or are microporous for an increased surface, though in EBA a fluidized bed is used. (wikipedia.org)
  • technique
  • Chromatography technique developed substantially as a result of the work of Archer John Porter Martin and Richard Laurence Millington Synge during the 1940s and 1950s, for which they won the 1952 Nobel Prize in Chemistry. (wikipedia.org)
  • liquid
  • It may be a liquid (LC and Capillary Electrochromatography (CEC)), a gas (GC), or a supercritical fluid (supercritical-fluid chromatography, SFC). (wikipedia.org)
  • Non-compacted particles remain mostly in the liquid called "supernatant" and can be removed from the vessel thereby separating the supernatant from the pellet. (wikipedia.org)
  • Agarose exhibits the phenomenon of thermal hysteresis in its liquid-to-gel transition, i.e. it gels and melts at different temperatures. (wikipedia.org)
  • chemically
  • It is not possible to separate chemically similar acids or bases using this simple method. (wikipedia.org)
  • A variety of chemically modified agaroses with different melting and gelling temperatures are available through chemical modifications. (wikipedia.org)
  • Temperature
  • Design and Preparation of a Novel Temperature-Responsive Ionic Gel. (patents.com)
  • Natural agarose contains uncharged methyl groups and the extent of methylation is directly proportional to the gelling temperature. (wikipedia.org)
  • Synthetic methylation however have the reverse effect, whereby increased methylation lowers the gelling temperature. (wikipedia.org)
  • commonly
  • The melting and gelling temperatures of agarose can be modified by chemical modifications, most commonly by hydroxyethylation, which reduces the number of intrastrand hydrogen bonds, resulting in lower melting and setting temperatures than standard agaroses. (wikipedia.org)
  • It is normally manufactured in a bead form and most commonly used for gel filtration columns. (wikipedia.org)
  • Columns
  • The purified antigens were radiolabeled and were passed through Sephadex G-200 or Bio-Gel A 1.5 columns to determine their m.w. (jimmunol.org)
  • The productivity of chromatography can be increased by running several columns at a time. (wikipedia.org)
  • A challenge that needs to be overcome in the future development of IC is the preparation of highly efficient monolithic ion-exchange columns and overcoming this challenge would be of great importance to the development of IC. (wikipedia.org)