Figure 1: Affinity chromatography of fumarase with the Ni2+-NTA-agarose column. Extract (9.9 mL) containing yeast (3.76g) in extraction buffer containing 0.1% Igapel CA-630 and protease inhibitors were pumped through Ni2+-NTA-agarose column. Fractions were collected by 1.5 mL portions by use of wash buffer (20.0 mL)‚ imidazole elution buffer (26.3 mL)‚ and wash buffer (10.0 mL)‚ again. Absorption readings were taken for all fractions with a Cary50 set at 280nm. The fumarase activity was determined
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Affinity chromatography technique is used to separate proteins found in a mixture of solution. Affinity chromatography uses the strong interaction between a given protein and its corresponding molecule. In today’s lab‚ affinity chromatography was used to purify L-lactate dehydrogenase‚ which contains histidine-tagged protein. The histidine- tagged protein forms a strong interaction with the Ni-NTA column due to the presence of nickel ions. Varying concentration of imidazole was added to the column
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Problem How can you tell pigment separation by using Chromatography? Objective To prepare a chromatogram‚ separate pigments in a leaf and interpret the chromatogram. Hypothesis If I am to put a chromatography paper into a solvent‚ then it would separate the pigments depending on their Rf value. I think the pigments will separate in this order: Chlorophyll A‚ Chlorophyll B‚ Carotene‚ and Xanthophyll. Introduction Chlorophyll is the molecule that absorbs sunlight and uses its energy
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Gas Chromatography Analysis of Product Mixtures Gas Chromatography Guidelines‚ Handout. Introduction Gas chromatography is a technique used to analyze chemical compounds that can be vaporized and separated in a gas phase column. Once separated‚ the analyzed substance is passed through a detector and data is obtained. The samples that we are going to analyze are: the EtOAc from Simple distillation‚ the Fraction 1‚ Fraction 2‚ and Fraction 3 from the Fractional Distillation. Experimental Procedure
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Information and Research 1. Chromatography is an analytical methor or technique that serves mainly as a tool for the examination‚ separating and identifying mixtures of chemical substances that are or can be coloured. 2.check the presence of any contamination in the manufactured compounds h as medicine‚ Contaminants in rainwater Analysis of narcotics Detection of substances in urine http://en.wikipedia.org/wiki/Paper_chromatography http://www.chemguide.co.uk/analysis/chromatography/paper.html http://chem
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In order to purify CelB2 protein bound to maltose-binding-protein‚ amylose affinity chromatography was performed. The amylose resin present in 20% ethanol was first diluted by adding 10 mL of 20 mM TrisHCl‚ pH 7.4‚ 0.2 M NaCl‚ 1 mM EDTA and centrifuged at 700 rpm for 5 minutes. After decanting the buffer‚ another 10 mL of TrisHCl‚ pH 7.4‚ 0.2 M NaCl‚ 1 mM EDTA was added to this resin solution and centrifuged at 700 rpm for another 5 minutes to further dilute the ethanol concentration in the resin
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The purpose of this lab was to understand that because substances have different properties‚ such as mass‚ that can be separated by chromatography. In our experiment‚ we chose 4 different color source to separate in water. We made sure that all the sources were water soluble because only polar substances will dissolve in water. We chose a black wet-erase marker‚ a red marker‚ a green marker‚ and mixed food coloring to test. The chromatography paper was split into 4 sections‚ about 2 cm above the
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Separation Of A Dye Mixture Using Chromatography ********* AP Chemistry (3rd Block) Fauquier High School 8/ 31/ 13 ABSTRACT: The Rf values of seven known color dyes and three unknown samples were observed in order to compare results and discover the hidden identities of the three samples. Three solvents‚ H2O‚ Isopropyl alcohol‚ and a 2% NaCl solution‚ were introduced to the dotted chromatography strips to show three different ratios for each dye on
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CHEM 2204 Chromatography Lab by wyk.wong » Fri Jul 11‚ 2014 10:25 am Results and Calculations Rf values Rf=(Distance moved by the spot (cm))/(Distance moved by the solvent front (cm)) Toluene: Rf=2 cm/3.8 cm=0.53 (Fluorenone) Rf=1.1 cm/3.8 cm=0.29 (Fluorene) Hexane: Rf=1.8 cm/2.2 cm=0.82 (Fluorene) Rf=0 cm/2.2 cm=0 (Fluorene Table 1: Experimental IR peaks compared to literature IR peaks for fluorenone Functional group Experimental peak (cm-1) Literature peak (cm-1) C-H 3010.5 3013
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Encyclopædia Britannica Paper chromatography is used to separate "dissolved chemical substances". An example of dissolved chemical substances could be the pigments that make up a specific color‚ for example in a marker. When a chromatography paper experiment is done on a black marker‚ the experiment tends to separate all the different pigments that make up the color. In this experiment‚ the substance investigated was a black water soluble Giotto marker pen. Paper chromatography has two phases‚ the
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