LDH Lab 1 - About 225 mg of protein and 5815 units of LDH...

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Dis 1N LDH Lab 1: First Progress Report In this first lab, eight grams of meat were taken and blended with a phosphate buffer to lyse the muscle cells, then centrifuged to separate the components based on density. The supernatant was poured through cheese cloth to filter out less dense fats and lipids; this resulting solution was called the crude extract. 40% and 60% ammonium sulfate fractionation was then performed on the extract in order to remove components differing in solubility from LDH, including fats and membrane components. The negative controls gave readings that stayed constant (the “no pyruvate” control had a slightly lower absorbance reading because of the absence of pyruvate).
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Unformatted text preview: About 225 mg of protein and 5815 units of LDH were found in the crude extract, giving a specific activity of 25.84. Following 40% ammonium sulfate fractionation, 185 mg of protein and 5531 units of LDH were measured, giving a specific activity of 29.90. Yield decreased by 5% and purification increased by 1.16 fold. No measurements need to be repeated for the next lab, since repeated measurements produced the same numbers with a small delta. Even though purification barely increased, these initial steps were important for removing components like fats and lipids, which would interfere with future assays....
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This note was uploaded on 05/10/2008 for the course BIO CHEM 153L taught by Professor Kim during the Spring '08 term at UCLA.

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