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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org/wiki/Chromatography http://ull.chemistry.uakron.edu/analytical/Chromatography/ http://orgchem.colorado.edu/hndbksupport/TLC/TLC.html this is for TLC – similar to paper http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/C/Chromatography_paper.html http://jchemed.chem.wisc.edu/JCESoft/Programs/CPL/Sample/modules/paprchrom/paprchromdesc.htm http://jchemed.chem.wisc.edu/JCESoft/Programs/CPL/Sample/modules/paprchrom/paprchromdesc.htm This site shows the colors of many of the food colorings
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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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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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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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Solubility as a Property of Matter A Lab of Chemicals‚ Chromatography‚ and Crime! Chemistry is a natural science that deals with the composition of matter and the changes it undergoes. At crime scenes‚ investigators often find unknown materials that need to be identified. If an unknown material is a mixture‚ an investigator may want to know one or two things about it: What are the ingredients of the mixture? Is the mixture found at the scene the same as a known mixture? A mixture is a collection
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Paper chromatography is an important separation technique that depends upon differences in how strongly the dyes are adsorbed onto the paper (stationary phase) and how soluble the dyes are in the developing solvent (mobile phase). In paper chromatography‚ a small amount of the mixture to be separated is placed close to the edge of a piece of paper. The edge of the paper is then immersed in a developing solution. As the developing solution ascends up the paper by capillary action‚ the. components
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Title:The Chromatography Process of Expo Markers Abstract:This experiment is to uncover whether or not the Expo Markers are homogeneous or heterogeneous mixtures. The experiment will conduct filter paper being slightly submerged in a container of water and inspected for pigmentation changes. In the experiment the pigments will rise up the filter paper once exposed to the water (solvent). After the experiment‚ it concludes that the mixture is a homogeneous mixture due to the chemical
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Liquid Chromatography Purpose: The purpose of this lab was to separate substances based on their polarity by using liquid chromatography. Data Table: Red Dye Blue Dye Run#1 Run#2 Run#3 Run#1 Run#2 Run#3 Start of Band(mL) 1.50 2.20 1.00 2.70 3.00 2.00 End of Band(mL) 2.70 3.00 2.00 6.40 5.50 6.00 Beaker Eluant Observations 1 H2O White powder 2 5%isopropyl Red powder 3 28%isopropyl Blue powder 4 70%isopropyl Oily residual Calculations: W = Vend – Vstart
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corresponding anthocyanidins using paper chromatography‚ and spectrophotometric analysis procedures. Based on experimental data the blueberries contained the most amount of anthocyanins at 1.92 x 10-6 g‚ followed by blackberries at 1.19 x 10-6 g‚ finally followed by raspberries at 5.84 x 10-7 g. The paper chromatography data also supports the idea that blueberries contained the most anthocyanidins‚ since it contained the most streaks on the chromatography paper. The main conclusion that can be
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