Unformatted text preview: Add the solution of proteins of known weight. Then add the eluent and collect fractions. To each fraction, add the blue protein dye. Analyze their absorbance using UV. Then add the hemoglobin to be characterized, and continue eluting until the hemoglobin has been eluted. Then test the absorbance of the hemoglobin and find the peak in its concentration. Compare the peaks of the first two proteins of known weight to the peak of hemoglobin. Figure 1. Based on the peaks of the two known molecules, BSA (66766 Daltons) and Lysozyme (17300 Daltons), it is estimated that the hemoglobin has a molecular weight of 61200 Daltons. The hemoglobin peaked at fraction 33 just one fraction later than BSA. So the estimation of 61200 is near the known weight of BSA. The literature weight for hemoglobin is 68000 Daltons. The experimental estimate is close to the literature weight indicating that is an efficient method for approximating molecular weight....
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- Spring '11
- Chromatography, gel permeation chromatography, Castleman Gel Permeation Chromatography