By Michail A. Alterman, Peter Hunziker
Amino Acid research (AAA) is an essential component of analytical biochemistry. In a comparatively little while, the range of AAA tools has developed dramatically with extra tools moving to using mass spectrometry (MS) as a detection procedure. one other new point is miniaturization. notwithstanding, most significantly, AAA these days might be considered within the context of Metabolomics as part of structures Biology. Amino Acid research: equipment and Protocols offers a huge spectrum of all on hand tools bearing in mind readers to settle on the strategy that the majority fits their specific laboratory set-up and analytical wishes. during this quantity, a reader can locate chapters describing common in addition to particular methods to the pattern guidance. a couple of chapters describe particular purposes of AAA in scientific chemistry in addition to in nutrition research, microbiology, marine biology, drug metabolism, even archeology. Separate chapters are dedicated to the appliance of AAA for protein quantitation and chiral AAA. Written within the hugely profitable equipment in Molecular Biology™ sequence layout, chapters comprise introductions to their respective issues, lists of the required fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and notes on troubleshooting and fending off recognized pitfalls. Authoritative and available, Amino Acid research: tools and Protocols offers the most important options that may be utilized throughout a number of disciplines via someone enthusiastic about biomedical learn or lifestyles sciences.
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Additional info for Amino Acid Analysis: Methods and Protocols (Methods in Molecular Biology, v828)
Divide the 10 min run time into time segments such that the minimum number of MRM transitions is observed per time window (see Note 9). If an Acquity UPLC system coupled to a Xevo TQ mass spectrometer is used, the optimized MRM method described in Table 1 can be used for the analysis of amino acids. 2. Metabolite Extraction from Arabidopsis Leaf Tissue 1. 5-mL microcentrifuge tube with screw caps/O-rings. Keep plant leaf tissue on dry ice before and after weighing. 2. 3 mm stainless steel beads to the microcentrifuge tube.
2. A comparison of detection limits and dynamic ranges obtained by AccQ·Tag-UPLC-ESI-MS/MS amino acid analysis using the Xevo TQ and TQD mass spectrometric detectors is given in Table 2. 63 × 10 (continued) 26 C. Salazar et al. 57 × 10 Dynamic range 4. Notes 1. Protect any unused reconstituted reagent solution from light and atmospheric water by wrapping the vial with aluminum foil, further sealing the cap with Parafilm, and storing the vial in a desiccator. Under these conditions, the reconstituted reagent last for up to 1 week.
Louis, MO) were used. Chemical purity of each amino acid standard was evaluated by the manufacturer and confirmed through in-house measurements (see Note 5). 3. 1. Calibrant and Sample Preparation 1. 1 mol/L hydrochloric acid. These stock solutions are stored at 5°C. 2. An internal standard solution is prepared by volumetrically blending the individual labeled amino acid stock solutions. The concentrations of the labeled amino acids in the internal standard solution should match as closely as possible the concentrations of amino acids in the hydrolysate of the protein or peptide solution being quantified.
Amino Acid Analysis: Methods and Protocols (Methods in Molecular Biology, v828) by Michail A. Alterman, Peter Hunziker