By Margaret I. Tyler (auth.), Catherine Cooper, Nicolle Packer, Keith Williams (eds.)
Amino acid research is primary in biotechnology, biomedical, and meals research laboratories. Amino Acid research Protocols constitutes an enormous choice of those imperative analytical thoughts, either vintage and state of the art, of excessive application for answering particular organic questions. universal equipment comprise these according to HPLC or gasoline chromatography separation and research after precolumn derivatization. New concepts in line with capillary electrophoresis separation, high-performance anion alternate chromatography, and mass spectrometry also are awarded. on the grounds that effects count seriously at the caliber of the pattern, such a lot members have committed a piece to pattern guidance, really to the gathering and garage of physically fluids. a brand new approach for desalting samples sooner than hydrolysis can be supplied. each one approach is defined in step by step aspect to make sure winning experimental effects, and comprises worthwhile notes on pitfalls to prevent, and diversifications that allow the how you can be used with various systems.
updated and hugely functional, Amino Acid research Protocols deals analytical and medical chemists, in addition to a huge diversity of organic and biomedical investigators, a wealthy compendium of laboratory instruments for the efficient research of either universal and unusual amino acids.
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And Chibata, I. (1976) Biosynthesis of norvaline, norleucine and homoisoleucine in Serratia marcescens. J. Biochem. 80, 333–339. 6. Tsai, L. , Lu, H. , Kenney, W. , Curless, C. , Klein, M. , et al. (1988) Control of misincorporation of de novo synthesized norleucine into recombinant interleukin–2 in E. coli. Biochem. Biophys. Res. Commun. 156, 733–739. 7. , Violand, B. , Dorward-King, E. , Workman, W. , Jung, P. , and Kane, J. F. (1989) Biosynthesis and incorporation into protein of norleucine by Escherichia coli.
5. After incubation, stop the reaction by adding 20 μL of 2 N HCl (see Note 5). Dry the reaction mixture under reduced pressure. 6. Store the dried samples at –20°C in dark until used (see Note 6). 7. Dissolve the dry sample in 1 mL of 50% (v/v) DMSO. 52 Kochhar et al. Fig. 2. Separation of 2,4-dinitrophenyl-5-L-alanine amide derivatives of L-amino acids by HPLC. A 20-μL aliquot from the amino acid standard mixture (standard H from Pierce Chemical Company) was derivatized with Marfey’s reagent.
Fig. 2. Separation of 2,4-dinitrophenyl-5-L-alanine amide derivatives of L-amino acids by HPLC. A 20-μL aliquot from the amino acid standard mixture (standard H from Pierce Chemical Company) was derivatized with Marfey’s reagent. 6 mm; 5 μm from Macherey-Nagel); sample, 100 pmol diastereomeric derivatives of 18 L-amino acids; solvent A, 13 mM trifluoroacetic acid plus 4% (v/v) tetrahydrofuran in water; solvent B, 50% (v/v) acetonitrile in solvent A; flow rate, 1 mL/min; detection at 340 nm; elution with a linear gradient of solvent B in solvent A (Table 1).