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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Chromatography serves mainly as a tool for the examination and separation of mixtures of chemical substances. Chromatography is using a flow of solvent or gas to cause the components of a mixture to migrate differently from a narrow starting point in a specific medium‚ in the case of this experiment‚ filter paper. It is used for the purification and isolation of various substances. There are two phases in chromatography: 1. Stationary Phase – a solid that does not move. In this experiment was the
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Thin Layer Chromatography Introduction Thin Layer Chromatography or TLC is a technique used as a separation and identification technique. There are many forms of chromatography‚ but one thing that remains constant throughout all of the types of chromatography is that there is a stationary phase and a mobile phase. In the case of TLC the stationary phase is the silica gel on the TLC tray. Procedure Chromatograph method is a method of separating mixtures of two or more compounds. Two phases
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Chromatography refers to a set of laboratory methods used in separating as well as purifying biomolecules. A variety of chromatography techniques exist‚ and all depend on the interaction between a stationary and a mobile state. Two types of chromatography methods were examined in this investigation. First‚ ion-exchange chromatography was used. This method separates ions and polar molecules based on their affinity to the ion exchanger [2]. Specifically‚ cation-exchange chromatography was performed
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Name: (e.g.‚ BioLab 3.4_A_Jones.doc) MDean Date: Graded Assignment Lab Report Answer the questions. When you are finished‚ submit this assignment to your teacher by the due date for full credit. (5 points) 1. Describe generally what happened to each spot of each type of ink. Which had the most pigments? Answer: The black ink went from a light blue to dark blue then to red. The red ink went from red to pink. It went the furthest out of the inks. The green ink went from dark blue to green and then
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Title: Thin-Layer Chromatography Objectives: Part1: 1. To learn the technique of TLC and the visualization of colourless components. 2. To identify an unknown drug by a TLC comparison with standard compounds. Part 2: To learn the separation technique by using Thin Layer Chromatography plate in separating a mixture of compounds into individual pure compound by using Spinach Leaf. Introduction: (i) General Concepts Chromatography is a common and powerful method used to separate and analyze complex
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Separation of Amino Acids by Cation Exchange Chromatography Introduction and Purpose: Amino acids are small biomolecules that have a carboxylic acid backbone in common‚ as well as an amino group attached to a saturated carbon. There are many amino acids‚ but there are 20 most commonly know amino acids. Amino acids are the fundamenta building blocks of other biomolecules like proteins and ezymes (Davidson‚ 2015). This experiment examined a mixture of 3 amino acids. The purpose of this experiment
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Column and Thin Layer Chromatography Beverly Abstract: Plant pigments were separated and concentrated from a crude spinach extract through the use of column chromatography and an eluatropic series of hexanes‚ hexane/acetone‚ and methanol. The pigments were analyzed using thin layer chromatography with a 30% ethyl acetate/hexane developing solvent. Introduction: Chromatography is a technique used to separate a mixture of two or more components based on
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possible to identify which peak correlates with which compound. Figure 2 contains a peak at around 500 and 700 nm which is a good indicator that that would-be chlorophyll b. This makes sense because this was the bottom band from the thin layer chromatography and chlorophyll b is the most polar compound. Chlorophyll B contains an aldehyde where chlorophyll A has a methyl group making it slightly more polar. Following that‚ figure 4 contains
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Task 3: How could Thin Layer Chromatography (TLC) have been used to prepare a pure sample of aspirin? The method of using thin layer chromatography is easy‚ quick and relatively cheap to carry out this makes it ideal for preparing pure samples of aspirin. Equipment Required: • Thin Layer Chromatography Paper • Ruler • Pencil • Solvent • Beaker • Micro Capillary • Access to a fume cupboard • UV light • Clingfilm • Individuals sample of aspirin • Pure sample of aspirin • Sample of salicylic acid Method:
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