By P. Alexander, R. J. Block

ISBN-10: 1483196259

ISBN-13: 9781483196251

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Additional info for Determination of the Size and Shape of Protein Molecules. A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides): Volume 3

Example text

PVW + RT\n Nw = μ„(Τ) + P°VW (6) The constant term μ^Τ) cancels out and it follows that nw = Ρ - i>° = - (RT/VJ In Nw (7) Uw = total osmotic pressure with a membrane permeable by water alone. In calculations of osmotic pressures less than 1000 m m Hg, it is customary to replace equation (7) by simpler equations, for example (8) a n d (9). n w = (RTjVw)Nx (8) Nx= 1 — Nw = molar fractions of solutes. (RTJVJ = 945,354 at temperature Τ of 273-16 or 0° a n d for volumes expressed in litres a n d pressures in millimetres of mercury of density 13-5955.

The tube Z), which consists of a short length of quill tube sealed on to capillary tube prevents protein from entering the capillary tube sealed below the bulb and thus from changing the density of buffer in the bulb. If the membrane be connected to the manometer at r o o m temperature, the initial pressure is set at a high level to compensate for contraction on cooling. The initial pressure is regulated by the distance to which the tube D is inserted in the rubber m o u n t E. The osmometer is then left to reach equilibrium at Γ and the pressure may be read on the next day.

More reproducible readings are obtained if osmotic pressure and capillarity readings are made after a falling rather than a rising pressure. A capillary tube of 1-3 m m bore, which contained 1-0% horse serum albumin in 0-02 M acetate buffer of p H 4-98, gave readings of 1-64 cm after 1 hr, 1-54 cm after 1 day and 1-46 cm 41 OSMOTIC PRESSURE after 9 days. Observations with a series of similar tubes showed a range of variation of 0-7 m m . A tube of 3-3 m m bore gave readings of 0-59 cm, 0-56 cm and 0-50 cm after the same time intervals.

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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


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