[CELL301] - Final Exam Guide - Ultimate 158 pages long Study Guide!
29 Nov 2016
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Tmrm is attracted to the highly negative mitochondrial matrix and fluoresces red when mmp is intact. Can be observed by microscopy or flow cytometry. Incubate cells with dye that fluoresces upon production of ros. Purpose: to determine whether a protein binds a specific region of dna. Analyzes labeled dna fragments on a gel after incubation with protein fractions. When proteins are bound to dna probes, there is a shift in molecular weight (and gel migration) Purpose: to identify a dna sequence in a gene enhancer that provides induction under certain conditions/proteins. When an inducible enhancer is placed in front of luciferase-encoding dna, and the conditions for transcription are met, identification via light emission. Extension of fragmented/laddered dna with fluorescent brdu (bromodeoxyuridine) Purpose: to isolate proteins found in the cytosol. Homogenization = physical rupture, leaves organelles intact (sonication, syringe) Lysis = addition of detergents - solubilizes organelles. Purpose: to purify proteins with different binding affinities to phosphocellulose.
