ANAT 365 Lecture Notes - Lecture 1: Lipid Raft, Lipid Bilayer, Differential Centrifugation
19 Jul 2016
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Heidi know stuff on slides, not what says. Sculpted events, lipids & proteins that bend them. Like fishing net across entire cell, holds cell together. Red fully saturated, straight chains pack tightly, high affinity for cholesterol, helps stabilize structure (don"t melt well) Blue unsaturated, kink mobile & fluid; would melt at high temp. At least one should be unsaturated for disordered domain. Add detergent at low temp; rafts don"t dissolve well, rigid. Add sugar, put at bottom of sucrose gradient. Centrifuge lipid structures float, proteins at bottom. Lipid rafts at top, collect fractions all in diff tube. Separate on sds page; sds denatures, apply current in gel, separates by size. Smaller goes to bottom, separates proteins in fraction. Nitrocellulose paper to blot, apply current to land on paper. Milk to block non-specific, antibody to label proteins, label w/secondary antibody to see. Can add antibody to megacluster megaraft, shifted up.
