Gel-Filtration Chromatography

Gel-Filtration Chromatography - Ashay Mehta Honors Chem....

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Ashay Mehta Honors Chem. 124 2/13/09 Gel-Filtration Chromatography Introduction - Chromatography is a method of separation in which components that are to be separated become distributed two phases. The two phases are the stationary phase bed and the other is the mobile phase. Separation occurs due to physical properties of the molecules, such as, charge and size. Gel-filtration chromatography is a method of separation due to the size of molecules. When separation occurs the small molecules pass though the pores in the beads that the large molecules cannot. Materials - 1. Column (15 mL) 2. Sephadex G-75 slurry 3. Elution buffer (20mm sodium phosphate, pH 7.0) 4. 16 x 100 mm test tubes 5. Sample mixture containing blue dextran, cytochrome c, and potassium chromate Procedure - 1. Pour a column so that the bed is about 13cm in height. Equilibrate the column in the elution buffer. 2. Add the mixture of blue dextran, cytochrome c, and Na 2 CrO 4 to the column and collect fractions. Observe whether these molecules are separated during the chromatographic
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This note was uploaded on 06/02/2010 for the course MGMT 121 taught by Professor Morozova during the Spring '10 term at NJIT.

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Gel-Filtration Chromatography - Ashay Mehta Honors Chem....

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