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
Premium DNA Bacteria Escherichia coli
Ink Chromatography Lab Background Information Paper chromatography is an analytical method that is used for separating and identifying mixtures of substances into their smaller parts. Paper chromatography works because the ink used contains several dyes (pigments) that when place on porous paper are dissolved in a solvent by capillary action. When the pigments (solutes) are dissolved in the solvent (water & alcohol mixture) they move through the paper at different rates depending on their
Premium Solubility Color Pigment
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
Free Chromatography Thin layer chromatography Solubility
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
Premium Chromatography Protein Analytical chemistry
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
Premium Thin layer chromatography Chromatography Solvent
Post Lab #4- Column Chromatography Organic Chem 3418-2 March 3‚ 2011 Theoretical Background- The fluorene and fluorenone mixture was separated by first dissolving the mixture in heptane. Since “like dissolves like”‚ fluorene dissolves with the non-polar heptane and the polar fluorenone dissolves in the polar ethyl acetate solvent. This phenomenon was illustrated in class before the experiment‚ when it was pointed out why water will not dissolve fluorene‚ fluorenone‚ or transstilbene
Premium Oxygen Chemistry Distillation
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
Premium Chromatography Gas chromatography Analytical chemistry
The purpose of column chromatography is to separate one compound from another by the differing polarities in the compound. This technique is important in organic chemistry because certain experiments may require the separation of compounds in solution to be used for chemical synthesis and/or analysis of a particular desired product by isolating it. Fluorene is to be separated from 9-fluorenone using the technique of column chromatography. The success of separation and purity is determined with analyzation
Premium Solvent Chemistry Solubility
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
Premium Cell Oxygen Chemistry
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
Premium Chromatography Analytical chemistry Thin layer chromatography