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Date: 13-12-2021
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Date: 21-11-2021
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Date: 16-10-2021
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Determining the amino acid composition of a polypeptide
The first step in determining the primary structure of a polypeptide is to identify and quantitate its constituent amino acids. A purified sample of the polypeptide to be analyzed is first hydrolyzed by strong acid at 110°C for 24 hours. This treatment cleaves the peptide bonds and releases the individual amino acids, which can be separated by cation-exchange chromatography. In this technique, a mixture of amino acids is applied to a column that contains a resin to which a negatively charged group is tightly attached. [Note: If the attached group is positively charged, the column becomes an anion-exchange column.] The amino acids bind to the column with different affinities, depending on their charges, hydrophobicity, and other characteristics. Each amino acid is sequentially released from the chromatography column by eluting with solutions of increasing ionic strength and pH (Fig. 1). The separated amino acids contained in the eluate from the column are quantitated by heating them with ninhydrin (a reagent that forms a purple compound with most amino acids, ammonia, and amines). The amount of each amino acid is determined spectrophotometrically by measuring the amount of light absorbed by the ninhydrin derivative. The analysis described above is performed using an amino acid analyzer, an automated machine whose components are depicted in Figure 1.
Figure 1. Determination of the amino acid composition of a polypeptide using an amino acid analyzer.
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في اليوم العالمي للصحة النفسية.. نصائح لتحسين مزاجك اليومي
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آلاف الأشخاص يحاولون الهروب من فلوريدا بسبب إعصار ميلتون
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بعد أسبوع من إطلاقها حملة الإغاثة للعوائل اللبنانية النازحة.. العتبة العلوية المقدسة تستضيف أكثر من (١٠٠٠) فرد مع تأمين الخدمات اللازمة
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