BIOC 3570 Study Guide - Midterm Guide: Freeze-Drying, Couette Flow, High-Performance Liquid Chromatography

• School: University of Guelph
• Department: Biochemistry
• Course: BIOC 3570
• Professor: David Josephy

Document Summary

Mass spec ion intensity vs m/z. Nmr abs vs. chemical shift (frequency) Seperation of molecules on basis of differences in retention by a stationary phase (matrix) through which a mobile phase (solvent) flows. A chromatogram is a plot of detector response vs time (elution volume) or distance. Seperation of molecules on the basis of charge characteristics (mobility in an electric field) An electropherogram is a plot of detector response as a function of time (elution volume) or distance (cid:498)henaed(cid:499) echnue identification) Chromatographic (or electrophoretic) separation followed by spectroscopic analysis (analyte. Polypeptides absorb light at = 280 nm. If you know the composition of protein, can calculate and c. Onl (cid:885) aa" cnbue b abon; trp is most important (aromatic aas) (cid:2870)8(cid:2868)(cid:4666)(cid:1839) (cid:2869)(cid:1855)(cid:1865) (cid:2869)=(cid:4666)#(cid:1872)(cid:1870) (cid:887)(cid:887)(cid:882)(cid:882)(cid:4667)+(cid:4666)#(cid:1872)(cid:1877) (cid:883)(cid:886)(cid:891)(cid:882)(cid:4667)+(cid:4666)#(cid:1855)(cid:1877)(cid:1871)(cid:1872)(cid:1866)(cid:1857) (cid:883)(cid:884)(cid:887)(cid:4667) Very rough rule: a280 protein concentration in mg/ml. Can provide evidence of presence of impurities. ~ 1 residue in 10 (trp/tyr) contributes to. Less sensitive than calorimetric assays which detect peptide backbone.