The use of metrizamide for stabilizing against convection in sedimentation equilibrium.
In meniscus depletion sedimentation equilibrium experiments, the solute concentration gradient in much of the cell is insufficient to stabilize against thermal convection. Experiments with spectrin at 35 degrees C led to sufficient convection that, particularly at low concentrations of solute, no apparent redistribution occurred. At low angular velocity, the density gradient provided by moderate concentrations of NaCl or additional sucrose is inadequate to overcome the convective disturbances. We have found that the use of low concentrations of metrizamide can overcome these problems. The higher molecular weight and lower partial specific volume of metrizamide vis-a-vis sucrose allow a significant and stabilizing density gradient even at angular velocities below 8000 rpm.
Ralston
G B
GB
Department of Biochemistry, University of Sydney, Australia.
Teller
D C
DC
Bukowski
T
T
D002499:Centrifugation, Density Gradient; D006863:Hydrogen-Ion Concentration; D008793:Metrizamide; D013049:Spectrin / Q000032:analysis; D013696:Temperature
D013049:Spectrin; D008793:Metrizamide