BIC472 - Assignment 5 - GFC.doc

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Seneca College
Chemical Laboratory Technician
Chemical Laboratory Technician BIC472
c A R L O B A S T I N U T T O

BIC472 – Assignment #5 – Gel Filtration Chromatography Professor: Carlo Bastianutto Student name: Daniel Katsafados Name of partner(s): Josh G Unknown: 1. (10 marks) Analyze your data including: • A graph of absorbance of the fractions versus the volume taken at that fraction. From this graph, determine the elution volume that gave the highest absorbance for each component (blue dextran, "mystery protein", and DNP- aspartate). • Your calculations of the Kdof the unknown protein • A standard graph of logMW versus K for tde data in Table 1. In this graph clearly show how you have determined the MW of the unknown protein. TABLE 1: K Vadues for Some Known Proteins on Sephadex G-75 Protein Molecular Weight K d (Daltons) Trypsin inhibitor (pancreas) 6500 0.70 Trypsin inhibitor (lima bean) 9000 0.60 Cytochrome-c 12400 0.50 α-Lactalbumin 15500 0.43 α-Chymotrypsin 22500 0.32 Carbonic anhydrase 30000 0.23 Ovalbumin 45000 0.12 K = (V – V ) = (15.5 – 5.0) = 0.53 d e 0 (V t V 0 (25 – 5.0) When extrapolating the graph the log molecular weight was 4.0 daltons, which when converted its 10, 000
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