By Cooper C., Packer N.
Proteome structures Ltd, North Ryde, Australia. Describes a number of amino acid research concepts and the way each one process can be utilized to respond to particular biologic questions.
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Extra resources for Amino Acid Analysis Protocols (Methods in Molecular Biology Vol 159)
1998) Use of the AQC Pre-column method for determination of protein composition and identification from PVDF blots. Presented at ABRF’98: From Genomes to Function — Technical Challenges of the Post-Genome Era, http://www. abrf. org/ABRF/ABRFMeetings/abrf98/rgrant. pdf. Amino Acid Analysis with Marfey’s Reagent 49 5 Amino Acid Analysis by High-Performance Liquid Chromatography after Derivatization with 1-Fluoro-2,4-dinitrophenyl-5-L-alanine Amide (Marfey’s Reagent) Sunil Kochhar, Barbara Mouratou, and Philipp Christen 1.
Mix 600 mL of acetonitrile with 400 mL of HPLC-grade water (Note: measure these volumes separately). 9 × 150 mm (see Note 7). 9 × 20 mm) packed with reversed phase packing, compatible with analytical column. Ternary gradient HPLC system equipped with an in-line degasser or sparging system, including column heater, autosampler, and dual monochromator fluorescence detector (see Notes 8–11). 3. 1. Hydrolysis 1. Use a pipet or syringe to place a protein or peptide sample in a 6 × 50-mm glass hydrolysis tube.
Corning, NY) (see Note 4). Reaction Vials (Waters Associates, Milford, MA). 2. Reagents 1. 2. 3. 4. 5. HPLC-grade water (see Note 5). Trifluoroacetic acid (TFA). Methanol. Hydrochloric acid (HCl). Phenol. 3. Reagents 1. 1 N HCl. 2. 1% phenol. 3. 1% TFA. 3. 1. Preparation of the Macrospin column (sample 50–150 µ L) 1. The macrospin columns should be prepared basically as suggested by the manufacturer (2), except that we have introduced several more washes to better equilibrate the columns. In brief, tap the column gently to recover the gel material at the bottom of the column.