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Gel Electrophoresis Lab Report

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In biochemistry it is very important to realize that the cells that make us who we are, are all made of specific proteins. These proteins need to be studied and analyzed extremely carefully to understand how certain proteins work in human physiology and in other organisms. In most cases it is important to separate one specific protein so you can better understand that protein’s function. One of the most common ways of separating these proteins is the use of gel electrophoresis. Gel electrophoresis separates proteins based on how big they are. The purpose of this lab was to isolate the protein in cultured E. coli that emits a green glow. The green glowing protein observed is known as Green Fluorescent Protein, or GFP for short. To separate the proteins inside the E. coli, gel electrophoresis and hydrophobic-interaction chromatography was used. Hydrophobic-interaction chromatography is used to separate proteins that are hydrophobic, GFP is hydrophobic therefore did interact with the hydrophobic-interaction chromatography. Knowing that GFP is hydrophobic, it is inferred that it is in the column. To know for sure that GFP was in the elution it was tested under UV light. If it glows green, then it means that GFP was located inside the sample. Once it was ensured that the GFP was in the sample, the electrophoresis …show more content…

Since the proteins should be separated on size only, these components are used. Sodium dodecyl sulfate is used to equalize the charge-to-mass ratio (charge density). SDS works by coating the proteins and disrupting structures to evenly distribute the negatively charged molecules, allowing isolation based on size only. Dithiothreitol breaks cysteine disulfide bonds, disrupting tertiary structures. This allows smooth movement throughout the gel. Last step in preparing the gel includes denaturing the protein at 95 degrees Celsius for 5

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