MBIO 3460 Lecture Notes - Lecture 12: Catalase, Microsome, Buoyancy
11 Jan 2018
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Document Summary
Understand the critical role centrifugation plays in cellular component isolation. Centrifugation is based on: sedimentation rate, differential centrifugation, buoyant density, density gradient centrifugation. Differential centrifugation: pellet formation, particles will sediment based largely on size, shape and density will also play a role, excellent for the crude separation of organelles and membranes, density gradient centrifugation is used for more precise isolations. Centrifugation: rotates sample at high speeds, greater speeds generate greater force, smaller particles within the pellet, speeds are measured in revolutions per minute (rpm) Also based on the: density of the medium, particle density, friction between the particles and the medium. The sample is layered onto the top of a stabilizing solution like sucrose. It can be a series of layered sucrose concentrations. The particles will band at specific zones based on mass: the band will lie above the next layer which is more dense than the particle.
