By P. Alexander, R. J. Block
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Extra info for Determination of the Size and Shape of Protein Molecules. A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides): Volume 3
The term k may be estimated from measurements of the rates made at two heights hx and h2 which differ by approximately 3 cm. (dhjdt) - (dhjdt) = -k(h2 - hd Precise measurements of these rates of movement can be made with a microscope as described in detail by Sorenson (1917). Exact measurements of k are not essential if the height alters very slowly. If, for example, dhjdt be less than 0-05 cm/day or 0-002 cm/hr, and k be greater than 0-10, the difference he — h, calculated by equation (33), is less than 0-02 cm.
Measurements of densities of protein solutions and of buffers together with formulae for calculating density increments have been given by Adair and Adair (1947). If the height h of the liquid in the manometer be not constant, but alters slowly with time, a small correction term may be estimated by applying equation (33). dh/dt = k(he - h) (33) h = observed height at time /; t = time in hours; dh/dt = rate in cm/hr; he equals height at equilibrium; k = rate in cm/hr when he — h — 1 cm. The term k may be estimated from measurements of the rates made at two heights hx and h2 which differ by approximately 3 cm.
The term pm is introduced so that the formula may be applicable to manometers containing toluene, tetrachlorethylene, or mercury, referred to later. In the osmometer shown in Fig. 2, y is equal to the distance between the level of dialysate W and the mid-point of the membrane, minus the length of the column of protein solution below the surface Wz in the device for determining capillarity corrections, shown in Fig. 3. In osmometers with a column of buffer above the protein solution, y is the distance between the mid-point of the membrane and the top of the column of protein solution.
Determination of the Size and Shape of Protein Molecules. A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides): Volume 3 by P. Alexander, R. J. Block