BIOC23H3 Lecture Notes - Lecture 2: Oxidative Phosphorylation, Cell Fractionation, Lysosome
17 Jan 2018
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Cell fractionation: cell fractionation is the process of separating specific layers/particles of different densities into groups, typically want to separate and purify different organelles. General procedure of cell fractionation: cell homogenization or opening of the cells, in cells with no cell wall you can change osmotic pressure of solution via hypo-osmotic buffer which will cause water to rush into cell, rupturing it. Want the buffer to mimic concentration of cytoplasm so organelles don"t rupture as well: cells with cell walls might require physical force via mortars, pestles, blenders, compression, ultrasonification. It is a preparative centrifugation where you separate and purify particles simply to collect them not to analyze its properties. Types of centrifuges: centrifuges have varying capacities and speeds, minifuge are the smallest, used mainly to quickly spin particles down to the bottom, 6,400 rpm max or 20000xg, microcentrifuge for spinning 1-2ml centrifuge tubes at speeds up to.
