DETERMINATION OF THE MOLECULAR WEIGHT AND PARTIAL SPECIFIC VOLUME OF HUMAN TRANSFERRIN BY MEANS OF A NEW TWO‐STAGE DENSITY GRADIENT EQUILIBRIUM METHOD

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Abstract
Using sodium diatrizoate as the gradient forming substance, the partial specific volume and molecular weight of human transferrin are determined by use of a new two‐stage density gradient equilibrium method. In the first stage of the method, equilibrium is reached with so low an angular velocity (24,630 rev./min) and column length (3.5 mm) that the resulting protein distribution corresponds to only a section of an isopycnic band. In the second stage of the method the initial protein concentration is determined by layering the salt solution on top of the protein solution in the centrifuge cell. The difference in refractive index between the two phases is studied as a function of time and appears to fall in a sequence of steps in one of the experiments. The difference between the first and second plateau of the graph is assumed to stem from a loss of water after the fluid injection and is used to correct the liquid density. The second plateau is used as a measure of the initial protein concentration in the first stage of the method.

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