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Unformatted text preview: Jason Kamin Bio 210-2 2/14/08 Lab #2 Protein Purification Introduction The main goal of this lab was to purify a mixture of proteins in order to analyze a certain protein, ICER. This sample of proteins was then put through a certain system of tests in order to test for the ICER. These tests included SDS-PAGE, western analysis and an electrophoretic mobility shift assay. All of these tests would test for the presence and purification of the ICER protein in a given sample and would provide different ways for analyzing the ICER protein and the processes used in its purification. Materials and Methods Part IA: Protein Purification: The different fractions of the proteins were to be made. This was done through the repeated use of centrifuge. In the first centrifuge done by the students the supernatant of the sample was saved and later used to make fraction 1, 2 and a pellet. This pellet was then saved and centrifuged five times, with various buffers at each step, in order to get a supernatant for fraction 3. These fractions would then be used in the subsequent steps of the lab. Part 1B: Polyacrylamide Gel Electrophoresis: In this part the gel electrophoresis set up was made and the student samples were loaded into assigned lanes of the apparatus. First the apparatus was set up by making sure the plates were even and screwed tight, and then the gel was loaded into the plates. The comb was then inserted into the gel in order to make the lanes, and after the gel polymerized, the lanes were ready to be filled with the student samples. With the samples loaded and the apparatus set up accordingly the only thing left was to wait for the electrophoresis to run and get the results....
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This note was uploaded on 02/16/2012 for the course BIOL_SCI 210-2 taught by Professor Somebody during the Winter '09 term at Northwestern.
- Winter '09