Chapter 3 Centrifugation
Chapter 3 Centrifugation Biochemistry and Molecular Biology (BMB) 3 1 Introduction 3 2 Basic Principle of sedimentation 3 3 Types, care and safety of centrifuges 3 4 Preparative centrifugation 3 5 Analytical centrifugation Analytical Biochemistry (AB) 3 4 3 Ultracentrifugation Koolman, Color Atlas of Biochemistry, 2nd edition
Ultracentrifugation: Probably the best downstream processing
Centrifugation KII ULTRACENTRIFUGATION Start volume 88L Rotor Volume Waste Volume 88L Gradient 60 Sucrose Flow Rate 15L/h Speed: 35 000 rpm Time 1 runs of 7h Gradient collected : Final Volume 600ml per run Concentrate 75x Yield 80 16
Analytical Ultracentrifugation in the Study of Protein Self
centrifugation, a 10 to 1000fold concentration range is typically observed, and a size-range of three orders of magnitude in molar mass can be covered in a single experiment Interacting components under study may have sizes ranging from peptides to very large multi-protein complexes In
AN OVERVIEW OF ANALYTICAL ULTRACENTRIFUGATION (AUC)
Analytical ultracentrifugation (AUC) is the most versatile, rigorous and accurate technology available for determining the molecular weight, hydrodynamic and thermodynamic properties of a protein or other macromolecule Although its history dates back nearly a century, AUC is:
Comparison of ultracentrifugation methods for concentration
with other viruses (Peng et al 2006; Gias et al 2008) After ultracentrifugation according to the first protocol, the supernatant was removed and the virus-containing pellet was resuspended by gentle agitation in 1 mL of TNE solution overnight at 4 °C After ultracentrifugation
Analytical Ultracentrifugation In Protein Science
equation (Teller et al 1979; van Holde et al 1998), in which the R o of the sphere is expressed as 1/3 4 3 π υ NA M, to obtain ( ) 1/3 4 3 6 1 π υ πη − υρ = A A sphere N M N M s (3) Substituting the values for all the constants (h for water at 20oC) yields Eq 4, the s value for a
PREPARATIVE DENSITY GRADIENT CENTRIFUGATIONS
Zone and isopycnic centrifugation methods can be combined, either as an analytical tool, for example to characterize the replicating complex of a viral genome (Magnusson et al , 1973), or as a preparative method for the purifi-
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