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Cooper C., Packer N.'s Amino Acid Analysis Protocols (Methods in Molecular Biology PDF

By Cooper C., Packer N.

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Constant boiling HCl. Phenol (crystalline or liquefied). 2. Derivatization 1. AQC dry powder. Store in a dry place, up to 6 mo in a desiccator. Available from Waters Corporation. 2. 8, with 5 mM calcium disodium ethylenediaminetetracetic acid (EDTA). 24 g of boric acid in a clean beaker. Add 100 mL of water, stir to dissolve. Add 187 mg of calcium disodium EDTA. 8 with sodium hydroxide (a solution made fresh from pellets). Use the best quality boric acid and EDTA available. 3. Acetonitrile for reagent dissolution.

Role of AAA in a Biotechnology Laboratory 25 Fig. 5. Analysis of carboxypeptidase digestion samples (expanded view). A standard mixture containing 1 nmol of each component was loaded. For clarity, only the early region of the chromatogram is provided to show the elution positions of Asn and Gln; the complete chromatogram is essentially the same as Fig. 8. Operating parameters are given in Table 6. Fig. 6. Amino sugar analysis. A hydrolysate from 1 nmol of a recombinant glycoprotein was loaded. Operating parameters are given in Table 7.

3. Marfey’s reagent from Pierce. Caution: Marfey’s reagent is a derivative of 1-fluoro-2,4-dinitrobenzene, a suspected carcinogen. Recommended precautions should be followed in its handling (15). 4. HPLC/spectroscopic grade triethylamine, methanol, acetone, and dimethyl sulfoxide (DMSO) from Fluka. 3. Chromatographic Analysis 1. Solvent delivery system: Any typical HPLC system available from a number of manufacturers can be used for resolution of derivatized amino acids. We have recently used HPLC systems from Bio-Rad (HRLC 800 equipped with an autoinjector AST 100), Hewlett Packard 1050 system and Waters system.

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