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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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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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 bottom edge and evenly spaced out. Each
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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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Gas chromatography (GC) is a type of chromatography that uses a carrier gas as the mobile phase and a column as the stationary phase. The sample is injected into the instrument and is heated until the sample‚ which includes both analyte and solvent‚ boils. The analyte must have
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Fractional Distillation & Gas Chromatography Exp. 1 Pre-Lab: 1) When two substances whose molecules are very similar from a liquid solution‚ the vapor pressure of the mixture related to vapor pressure of the pure substance. Also it could be defined as a two liquid are ideal solution when they don’t react with each other and they make no association. 2) Are a mixture of at least two different liquid‚ and known also as a mixture of two or more liquid in such away that its component
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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 yellow. Out of the 3 inks‚ the green ink had the most pigments. (5 points) 2. Compare the effectiveness of each of the four kinds of solvents. Answer: Water will move the ink
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experiment is to identify an unknown proprietary drug using thin-layer chromatography. The unknown’s behavior in thin-layer chromatography will be compared with that of its possible component analgesics. The possible unknowns and their analgesic ingredients will be Anacin (aspirin‚ caffeine)‚ Excedrin (acetaminophen‚ caffeine‚ aspirin)‚ Motrin (ibuprofen)‚ and Tylenol (acetaminophen). Introduction Thin-layer Chromatography(TLC) was the method used to figure out and identify the drug. TLC is done
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Purification of Recombinant Green Fluorescent Protein (rGFP) From E. coli strain‚ BL21(DE3)‚ Using Ni2+-Agarose Affinity Chromatography Abstract: The purpose of these series of experiments was to express and purify recombinant Green Fluorescent Protein (rGFP) from the E. coli strain‚ BL21(DE3) by beginning with its purification via a Ni2+-agarose affinity chromatography column. The His6 tag of the rGFP bound to the Ni2+-agarose column and washes and elutions were obtained‚ with elution 3 containing
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Mobile liquid phase is always passed stationary phase during chromatography. Developed TLC plate is then observed under short wave ultraviolet light‚ where most TLC plates contain fluorescent mixed with silica gel that allow to glow green color. Compounds will absorb the UV light and appear as dark spot against the green color TLC plate. In a column chromatography‚ cotton and sand is placed in purpose of holding silica gel and prevent leakage of adsorbent particles
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