protein was filtered out of the cell and column chromatography was used as a means to isolate the protein by taking advantage of its hydrophobic properties. This was performed by washing the column containing the GFP protein amongst other proteins with buffers varying in salt concentrations. As expected‚ when a buffer of high salt concentration was used (binding buffer) the protein remained in the column. This was evidenced by the green glow of the column coupled with the non-glowing wash remnants
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antioxidant. Lycopene is found in various fruits and vegetables. The purpose of this experiment is the isolation of Lycopene from tomato paste. This is done using liquid/solid extraction and chromatography. Once the Lycopene is isolated‚ IR spectroscopy will determine its percentage actually obtained by chromatography. Procedure: A massed sample of 1.012g of tomato paste was placed in a 125mL Erlenmeyer flask. To the flask‚ 5mL of 50:50 hexane-acetone was added into the flask. After the 50:50
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injection loop is pulled down to create a high pressure pump that pushes the sample into the mobile phase of the instrument. This mobile phase‚ which for this experiment will consist of water and methanol‚ transports the samples toward the packed column where it will leave the mobile phase and enter the stationary phase. In this stationary phase‚ the sample is fixed in place for the amount of time it takes for the sample to elute back to the mobile phase. This stage identifies and quantifies the
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A0133901R 1. Aim 1. To isolate chlorophyll and beta carotene from plant leaves using column chromatography. 2. To qualitatively characterise the pigments with UV-vis spectroscopy and TLC. 2. Results and discussion Isolation of beta carotene and chlorophylls by column chromatography Upon the loading of S1 (the extract of the plant leaves in hexane)‚ a yellow band appeared at the top part of the silica column immediately after the solvent level descended to just above the sand layer. This yellow
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the column before we could get an absorbance rate of 0.1‚ which means that there was much of our purer LDH sample that we were unable to collect and use. Another thing to note is that our values for the affinity chromatography step were almost the same as the amount for the size exclusion chromatography step. The total activity was 96.50U and the percent yield was 29.76% for the size exclusion. The lack of purity and low recovery of our size exclusion chromatography could be due to the column beads
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L-Lactate Dehydrogenase By Affinity Chromatography on Cibacron-Blue Sepharose David Alexander 10-15-2014 Dr. Black Chem 4135.001 Abstract: Like the previous experiments‚ the ultimate goal of this lab was to purify the enzyme sample. However‚ this is the last lab for purification and high level techniques of purification were employed to achieve this. Dialysis was used first‚ lowering the small-molecule concentration within the sample. Finally Affinity Chromatography on a Cibacron blue Sepharose stationary
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Analysis of Lipid through Two-Dimensional Thin Layer Chromatography Myca Pua‚ Ramon Ramos‚ Amanda Redilas‚ Kimleigh Reyes‚ Nathaniel Sim and Clara Tamondong Group 9 2F Medical Technology Biochemistry Laboratory ABSTRACT Lipids constitute a group of naturally occurring molecules that include fats‚ waxes‚ sterols‚ fat-soluble vitamins such as vitamins A‚ D‚ E‚ and K‚ monoglycerides‚ diglycerides‚ triglycerides‚ phospholipids‚ and others. In this experiment lipid was extracted from the egg yolk
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Performance Liquid Chromatography but is also referred to as High Pressure Liquid Chromatography. It is used for separating mixtures either to analyse the mixture or to separate a required product from others in a reaction mixture. It can also be used to find the relative amounts of different components in a mixture. HPLC works along the same lines as paper chromatography. In paper chromatography a liquid (mobile phase) moves past a solid (stationary phase). In paper chromatography the stationary phase
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support‚ silica gel‚ which acts as a Lewis acid catalyst to facilitate the reaction. The reaction forms a porphyrinogen‚ which is then oxidized to the porphyrin product by atmospheric oxygen. Column chromatography is performed for the isolation and purification of tetraphenlyporphin‚ and the thin layer chromatography for analysis.It was concluded that the renention factor(Rf) of the 5‚10‚15‚20- Tetraphenylporphyrin with a percent yield of 61% Introduction: In this experiment 5‚10‚15‚20‚ tetraphenylporphyrin
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protein capture by Protein A affinity chromatography helps to remove a large majority of the impurities whicn include viruses and media components. At Stage 3‚ prior to cation exchange chromatography‚ the purified material in the eluted stream is then subjected to freezing‚ thawing and pooling. Stage 4 involves the Solvent/Detergent (S/D) viral inactivation which is the first dedicated viral clearance step. At Stage 5‚ the product undergoes cation exchange chromatography which aids in removing the S/D reagents
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