BIOS 10161 Study Guide - Quiz Guide: Bromophenol Blue, Ionic Strength, Agarose Gel Electrophoresis
3 Sep 2016
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Electrophoresis: movement of charged molecules under the influence of an electric field. Effective for protein mw determination; dependent on size. ; dependent on composition of aa residues and ph of chamber buffer// isoelectric point. Sds (hydrocarbon chain with ive sulfate group) binds strongly to proteins. Functional groups are ionized or non-ionized depending on pi and buffer; 10/4 rule. Reducing agent: b mercaptoethanol breaks disulfide bridges and completely dissociates proteins. Charge depends on ph of buffer used isoelectric point of proteins. Separation by differences in isoelectric points of proteins. All samples contain sucrose/glycerol to create density > density of electrode buffer sample can sink into well. Tracking dye (bromophenol blue- bpb) migrates ahead of smallest proteins toward anode. Tracking dye (bromophenol blue- bpb) w/ strong ive charge migrates ahead of most negative// fastest proteins. Stained for visibility and fixation (prevents protein diffusion)
