Download e-book for kindle: A Laboratory Manual of Analytical Methods of Protein by P. Alexander, H. P. Lundgren

By P. Alexander, H. P. Lundgren

ISBN-10: 0080113982

ISBN-13: 9780080113982

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Additional resources for A Laboratory Manual of Analytical Methods of Protein Chemistry. Volume 4

Sample text

Using the above precautions, MeHgl and the other reagents mentioned have been in constant use in the reviewer's laboratory by 6 or 8 workers for nearly 10 years without any reported ill effects. MeHgl (5 x 10~4M)—Solution A: This solution is required for measuring the rate and final extent of reaction of —SH groups with excess MeHgl. Since dénaturants are absent, any changes in biological or physicochemical properties of a protein treated with this solution may be attributed to reaction of the —SH groups alone.

When insoluble samples are used, the flasks should be examined after the first hour or two to ensure complete wetting and immersion in the mercurial solution. It is sometimes necessary to apply suction to remove adherent air bubbles. After reaction, the solution is decanted non-quantitatively into the polarographic cell* so * More complete details of the operation of the polarographic cell and of precautions to be observed, are to be found in method (b). ESTIMATION OF THIOL AND DISULPHIDE GROUPS 39 that the dropping mercury capillary is just dipping into the solution.

In certain conformational studies on proteins and enzymes, the conversion of —SS— to —SSOJ groups by treatment with sulphite in the presence of oxidizing agents has become a useful technique for splitting —SS— bonds (Swan, 1957; Bailey, 1957; Péchère et al, 1958; Weil and Seibles, 1959). It is often important to know whether —SS— fission has been complete. However hydrolysis of the S-sulpho protein results in partial conversion of —SSO " to —SH groups accompanied by some destruction. In some cases destruction is complete.

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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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