BIOL 1300 Midterm: Midterm 1 Review
Document Summary
To know: advantages, disadvantages, how do they work, how they were applied in papers. To know: all the figures from the research articles - be able to interpret every single aspect of every single figure without any context :) In vitro binding/pull down assay: how it works: add tag (gst, 6x his, etc. ) to protein to make a purified fusion protein. -> add tagged protein to lysate --> see what is pulled down with it/what molecules it binds. Can then digest via trypsin, ms, sequence to determine identity of molecule: advantages: simple, compatible with downstream analysis methods, disadvantages: requires significant amount of recombinant dna manipulation, not suitable if a third party mediates the interactions. Blue/clear native page: blue how it works: identifies complexes that stay together - coomassie anionic blue die stains arg residues, stabilizes protein complexes --> separated by native page --
