Download PDF by P. Alexander, H. P. Lundgren: A Laboratory Manual of Analytical Methods of Protein

By P. Alexander, H. P. Lundgren

ISBN-10: 0080113982

ISBN-13: 9780080113982

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Extra info for A Laboratory Manual of Analytical Methods of Protein Chemistry. Volume 4

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Other radiochemical techniques will be mentioned in this section; although they are not yet widely used, they offer great promise for precise and automated analyses at the micro and submicro level. Some of the pitfalls to be encountered in analysing —SH and —SS— groups in proteins after they have been hydrolysed will now be enumerated. B. Analysis after Hydrolysis 1. Hydrolysates of Proteins Although alkaline hydrolysis is still sometimes used prior to (Ogura et al, 1963; Kusuhara and Knox, 1962) or during (Wronski, 1963c) —SH and —SS— estimation, its use is to be deprecated.

The other end of the KC1 bridge dips into compartment (B) of the calomel electrode (Fig. 4 (b)). The mercury reservoir is raised to start the electrode dropping and the capillary tapped a few times to ensure that no N 2 bubbles are occluded at the tip. The polarograph is set to apply a potential of —0-6 V (simple thiols) or — 0*7 V (proteins) to the dropping mercury electrode, relative to the calomel electrode, and the current (microamps) is recorded. After each addition of titrant (0-1 ml) tap (H) is closed to force the aliquot into the cell, and the side-arm washed out by closing and opening the tap as already described.

The process is illustrated in Fig. 9 for the case of a fibrous keratin. ) FIG. 9. Determination of thiol groups in fibrous keratins. Current-voltage curves for MeHgl (5 x 10 - 4 M) in tris buffer (pH 7-2) in the presence of KC1 and gelatin (ca. 0-01%): (a) before reaction with keratin; (b) after reacting MeHgl (10 ml) with keratin (44-4 mg) for 24 hr at 20°C. The reduction in the height of the wave corresponds to a value of 37 micromoles of —SH per g of keratin. There is no additional uptake even after 88 hr (Leach, 1960b).

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A Laboratory Manual of Analytical Methods of Protein Chemistry. Volume 4 by P. Alexander, H. P. Lundgren


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