Amino acids analysis Protocols by Catherine Cooper

By Catherine Cooper

A suite of vintage and state-of-the-art strategies of excessive software in answering particular organic questions about amino acids. universal equipment contain these in accordance with HPLC or fuel chromatography separation and research after precolumn derivatization. New thoughts in accordance with capillary electrophoresis separation, high-performance anion alternate chromatography, and mass spectrometry also are awarded. each one technique is defined in step by step aspect to make sure profitable experimental effects and emphasizes pattern practise, fairly the gathering and garage of physically fluids. updated and hugely useful, Amino Acid research Protocols bargains analytical and scientific chemists, in addition to a vast diversity of organic and biomedical investigators, a wealthy compendium of laboratory instruments for the effective research of either universal and unusual amino acids.

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2. Chromatographic eluents: 18 megohm water from a Milli-Q (Millipore, MA), 50% sodium hydroxide (UN1824, Certified Grade, Fisher, Pittsburgh, PA), anhydrous sodium acetate (71179, Microselect Grade, Fluka, Milwaukee, MI). 3. Diluents and standards: norleucine (N1398, Sigma), sodium azide (S 8032, Sigma), Standard Reference Material Amino Acids (SRM 2389 NIST, Gaithersburg, MD), cysteic acid (C7630, Sigma), methionine sulfone (M0751, Sigma), methionine sulfoxide (M1001, Sigma), Mixture of Six Carbohydrates (P/N 43162, Dionex).

The PVDF membrane should not be allowed to dry between aliquots. 15. Based on a recent ABRF study on amino acid analysis (8), samples bound to PVDF membranes tend to have higher error rates (20%) than those analyzed free in solution (12%). These error rates are averages for the study. References 1. Dupont, D. , Keim, P. , Chui, A. , and Wilson, K. J. (1989) A comprehensive approach to amino acid analysis, in Techniques in Protein Chemistry (Hugli, T. ). Academic, London and New York, pp. 284–294.

04 g of anhydrous sodium acetate in approx 750 mL of pure deionized water. Seal the container during the dissolution step. 2-µm Nylon filter. We recommend using sterile Nalgene vacuum filtration units for this step (see above). Transfer the filtered sodium acetate eluent into the eluent bottle E3 of the DX-500, using the same procedure as for the sodium hydroxide. Diluent solutions containing sodium azide: Prepare 1 or 2 L of an azide diluent solution containing 20 mg NaN3/L. Use the solution without norleucine whenever an internal standard is not required.

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