BIOB10H3 Study Guide - Final Guide: Mass Spectrometry, Fluorophore, Diffraction
Document Summary
Step 1: protein purification go to revision package 6. Now we can start to answer some of these scientific questions: primary protein sequence: mass spectrometry. The sequence determining the order of its aa. Protein purified from cell extract may be cut out of a 2d gel. Protein is broken into smaller pieces (peptides) using proteases (e. g. trypsin) These peptides are dried onto a metal plate & heated by a laser. Makes peptides electrically charged (ionized) with +ve ions knocking one or more e- off. Fired from the plate as a gas through a magnetic field within a machine called a mass spectrometer. Peptide ions separate from each other based on their mass and charge; Smaller ions travel faster than larger ions; highly charge ones move faster also. Separate according to their mass to charge ratio = m/z. Ions hit a detector o(cid:374) o(cid:374)e e(cid:374)d (cid:449)hi(cid:272)h plots the (cid:858)sig(cid:374)atures(cid:859) of the peptides based on the time it takes to reach it.
