Chemical Laboratory Technician BIC472 Lecture Notes - Trypsin Inhibitor, Carbonic Anhydrase, Phaseolus Lunatus
21 Mar 2013
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Bic472 assignment #5 gel filtration chromatography. 1. (10 marks) analyze your data including: a graph of absorbance of the fractions versus the volume taken at that fraction. In this graph clearly show how you have determined the mw of the unknown protein. Table 1: kd values for some known proteins on sephadex g-75. Kd = (ve v0) = (15. 5 5. 0) = 0. 53 (vt v0) (25 5. 0) When extrapolating the graph the log molecular weight was 4. 0 daltons, which when converted its 10, 000 daltons. 2. (2 marks) why did you observe two peaks when you measure the absorbance at. 3. (4 marks) you are asked to purify the protein bic472 from a sample containing a mixture of proteins. Dnp-aspartate to your mixture, you run it into a sephadex-g75 column and collect 20 fractions of 0. 5ml. Blu-dextran elutes in fraction #3, for a volume of. 4. 5ml, while dnp-aspartate elutes in fraction #18, for a volume of 12ml.
