Gel filtration chromatography on Sephadex G-50 The crude broth obtained after fermentation was subjected to ammonium sulphate precipitation at 70% (w/v). The pellet so obtained was resuspended in cold saline (2 ml) and dialysed. The dialysed enzyme was loaded onto a column of Sephadex G-50 (120 cm × 1.0 cm) equilibrated with 10 mM Tris-HCl buffer‚ pH 8. The column was eluted at a flow rate of 1 ml / 6 min. The elution profile of gel filtration chromatography is shown in the (Fig: 1). The fractions
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Title: Liquid Chromatography Author: Gloria Contreras Lab Partner: Jose Montanez Instructor: Teresa Potter Date Work Performed: January 13‚ 2015 Date Submitted: January 20‚ 2015 Abstract: In this lab‚ liquid chromatography is used to separate the Red 40 and Blue 1 dyes inside of grape flavored Kool-Aid. It was determined that the 5% isopropanol will remove the Red 40 dye from the stationary phase. The 28% isopropanol will remove the Blue 1 dye from the stationary phase. The more
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purified by flash chromatography and analyzed by IR‚ 1H NMR and 13C NMR. The synthesis of piperonylontitrile was successful. There was a yield of a pale yellow crystal of 36%. The average recovery is 51% with there being a range from 0-94 %1. A reason for a smaller yield than notable reported could have been due to transfer of the product from glassware to glassware. Some may have been left behind. Another reasoning for a smaller yield could be due to the fact that as soon as the column was ran the product
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the E.coli strain using Ni2+ agarose affinity chromatography technology Abstract The purpose of this experiment was to express and purify the his6-tagged recombinant form of GFP (rGFP) from the organism E.coli using Ni2+ agarose affinity chromatography. The expression of rGFP was confirmed qualitatively using the UV light and was expressed in the E.coli strain BL21 (DE3) (-- removed HTML --) (-- removed HTML --) . Ni2+ agarose affinity chromatography was used to purify the crude extract and different
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Ion Exchange Chromatography Discussion: The first exercise preformed in this lab was ion exchange chromatography. The purpose of this experiment is to separate molecules based on their differences in charge. Since it is based on charge the amino acids in the cation exchange column‚ if negatively charged‚ flow through the column first because they don’t want to bind to the sodium ions. The positively charged ions will elute last at the highest ph because they bind to the negatively charged beads
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use Gas Chromatography to distinguish between two enantiomers of carvone from caraway oil and spearment oil and to find the 2 carvone’s optical activity as well as percent carvone in spearment and caraway oil. It was found that S-carvone had an optical activity of 0.0047 and R-carvone had an optical activity of 0.516 and that spearment oil is 59% carvone and caraway oil is 100% carvone. Backround: Gas Chromatography separates organic samples much in the same way as column chromatography. The only
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The knowledge of chemical analysis today has become important not only for scientists in their research but in fact bears influence in our daily routine as well. To recognize and classify these uses‚ we have to first understand‚ what is chemical analysis?Chemical AnalysisChemical Analysis is a body of procedures and techniques used to identify and quantify the chemical composition of a sample of substance.1 It may be said as a collection of all the techniques required to obtain any chemical information
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dyes‚ the resolution should be greater than one. What was the value you calculated? Did the two dyes overlap as they emerged from the column‚ or was the separation a good one? The resolution calculated was .778 (see calculations)‚ which made it difficult to perceive where exactly one dye began and the other ended. Thus‚ when the dyes were emerging from the column‚ they mixed and formed a blend of the two colors to form a light purple color‚ which made it difficult to ascertain where the red ended
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solubility of a solute (a dissolved substance) in a solvent (the dissolving medium) is the most important chemical principle underlying three major techniques you will study in the organic chemistry laboratory: crystallization‚ extraction‚ and chromatography. In this experiment on solubility you will gain an understanding of the structural features of a substance that determine its solubility in various solvents. This understanding will help you to predict solubility behavior and to understand the
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Title: Competing Nucleophiles (Exp 24‚ pp 211-221‚ pp 808-823‚ pp 836-842) Purpose: The purpose of this experiment is to determine the nucleophilic strength of chloride and bromide ions as it reacts with 1-butanol (n-butyl) and 2-methyl-2-propanol (t-butyl alcohol) under SN1 and SN2 conditions. Method: 40 g of ice and approximately 30 ml of sulfuric acid is cautiously added to a 100 mL beaker respectively. Weigh 7.6 g of ammonium chloride and 14.0 g of ammonium bromide and place it in
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