such as activity and protein assay were employed to determine the presence and purity of LDH. The cells were initially disrupted and proteins were solubilized. LDH was purified from the ammonium sulfate precipitated protein mixture by affinity chromatography and its activity was studied by spectrophotometric determination of NADH at 340 nm. From Pierce BCA assay of crude homogenate‚ initial protein concentration was shown to be 100 mg/ml. The final protein concentration of the pooled affinity sample
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inhibiting enzymatic reactions involving para-aminobenzoic acid (PABA). PABA is needed in enzymatic reactions that produce folic acid which acts as a coenzyme in the synthesis of purine‚ pyrimidine and other amino acids. In part C‚ Fluorene was dissolved in three solvents. Fluorene(C13H10) is a polycyclic aromatic hydrocarbon. It forms white crystals that exhibit a characteristic‚ aromatic odor similar to that of naphthalene. It is combustible. It has a violet fluorescence‚ hence its name. For commercial
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binds the nucleoside inhibitor site. When it binds this site‚ it stabilizes the inactive T state and blocks the catalytic site which needs to be open for enzyme activity to occur. The glycogen phosphorylase b was purified with hydrophobic column chromatography and the concentration was determined with a Bradford Assay. The kinetics of glycogen phosphorylase b were studied by finding a molar extinction coefficient‚ 0.1617 mM cm-1 A-1‚ for
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Chromatography of Commercial Analgesics Chromatography of Commercial Analgesics CHEMISTRY 200L EXPT 4 PAGE 8 - 11 CHEMISTRY 200L EXPT 4 PAGE 8 - 11 Janna Vernice R. Villalon*‚ Christian V. Villanueva‚ Cyd Vincent L. Zamora Department of Chemistry‚ College of Science *Corresponding author; e-mail: janna.villalon@rocketmail.com Abstract In analyzing the chromatography of analgesics‚ thin layer chromatography (TLC) was used. A very thin (micron) film of silica is coated on a glass
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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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