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Size Exclusion Chromatography- Protein Separation Polyacrylamide Gel Electrophoresis - Lab Report Example

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The absorbance of each fraction at 200nm was determined and plotted as shown in figure 1 below. Even though the resolution between the…
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Size Exclusion Chromatography- Protein Separation Polyacrylamide Gel Electrophoresis
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"Size Exclusion Chromatography- Protein Separation Polyacrylamide Gel Electrophoresis"

Download file to see previous pages These standards were used to plot the calibration curve.
The retention times, which is equivalent to elution volume (Ve) of each protein was plotted as a function of the log 10 of the molecular weight of the standards as shown in figure two. This calibration curve was used to estimate the relative molecular weights of the unknown samples.
The Value for X and Y in this function is equivalent to elution volume (Ve) and Log10 of the molecular weight. Substituting Ve with the retention time value, the Log M and molecular weights of the unknown samples were determined as shown on table three below.
Size exclusion chromatography is a HPLC technique that separates molecules based on their size or molecular weight (Bernd, 2). Using this technique the molecular weight of five unknown protein molecules was estimated as shown in table three. However, the validation assay of the technique did not give expected resolution, limits the validity of the results. Clear and distinct peaks on this graph depict a high resolution of the assay and thus credible results. A number of factors; such as column length, flow rate and technical errors may have influenced the poor resolution of the peaks in the UV protein absorption profile (Hong and Fountain, 5-6). Further optimization experiments may be necessary to overcome these errors and achieve a good ...Download file to see next pagesRead More
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