Amino Acid Mixes
To aid research development and application of amino acids, CIL has formulated various mixtures of stable isotope-labeled (and unlabeled) amino acids. These include mixes of canonical and branched-chain amino acids (MSK-CAA and NSK-BCAA), a mix of rare or unnatural noncanonical amino acids (MSK-NCAA), and mixes of amino acids/derivatives (e.g., NSK-A, NSK-AA3). All mixes are well characterized for use in quality control and qualification/quantification studies.
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Amino Acid Mixtures
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❛❛I use several of CIL’s metabolomics mixes in my LC-MS analysis. They make quantitative metabolomic work convenient with a single internal standard mix spike and provide the corresponding unlabeled mix for absolute quantitation. The ready mixes save time from the tedious task of making up individual solutions of each analyte for quantitation. I find the different mixes applicable to a variety of analyses in complicated matrices that provide metabolic insight to my studies.❜❜
– Andrew Downey, Senior Scientist, Axcella Health Inc. (USA)
Frequently Asked Questions
In what form are the amino acid mixes? The MSK-CAA mixes (labeled, MSK-CAA-1; unlabeled, MSK-CAA-US-1) and MSK-NCAA mixes (labeled, MSK-NCAA-1; unlabeled, MSK-NCAA-US-1) are dried down, while the MSK-A2 mixes (labeled, MSK-A2-1.2; unlabeled, MSK-A2-US-1.2) are supplied as a 1.2 mL solution (in 0.1 M HCl).
What are the metabolites and concentrations in the described amino acid mixes? Please refer to our Stable Isotope-Labeled Mixtures, Sets, and Kits catalog for the compositions. Note that the metabolites and concentrations in a given unlabeled mix match their labeled counterpart (e.g., MSK-CAA-US-1 is equivalent to MSK-CAA-1, with the only difference being the stable isotope labeling).
In what matrices has MSK-A2-1.2 been applied? This mix has been spiked (after dilution from its concentrated stock) into a range of human-based matrices (e.g., cell, tissue, plasma, and serum). An example set of references are listed below.
What is the shelf life of the amino acid mixes? The recommended retest for all three MSK amino acid mixes is two years if stored in a refrigerator (-5 to 5°C, protected from light). Upon reconstitution, the amino acids in MSK-CAA mixes (labeled, MSK-CAA-1; unlabeled, MSK-CAA-US-1) and MSK-NCAA mixes (labeled, MSK-NCAA-1; unlabeled, MSK-NCAA-US-1) are stable in the refrigerator (-5 to 5°C, protected from light) for four weeks.
How do the MSK-CAA mixes differ from MSK-A2 mixes? Relative to MSK-A2 mixes (labeled, MSK-A2-1.2; unlabeled, MSK-A2-US-1.2), MSK-CAA mixes (labeled, MSK-CAA-1; unlabeled, MSK-CAA-US-1) contain three additional amino acids (L-Asn, -Gln, and -Trp) and are dried down. This relates to the reconstitution condition and volume, which may impact the choice of sample preparative method and analysis technique.
What is an appropriate solvent for solubilizing the MSK-CAA and -NCAA mixes? Suitable examples of dissolution solvents are water or water/methanol. It is imperative that the CAA mix is not reconstituted in 0.1 M HCl, as L-Asn and L-Gln are known to be unstable in acid.
Does CIL offer the ability to make custom mixes? Yes, we do have the ability to customize. We would first review feasibility and then provide a quotation on your specific mix. To start this process, please provide the necessary details on this Custom Mix Request form or contact your local sales representative.
Example References
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McNabney, D.W.G.; Mangal, V.; Kirkwood, A.E.; et al. 2023. Phytoplankton metabolite profiles from two Lake Ontario areas of concern reveal differences associated with taxonomic community composition. Sci Total Environ, 871, 162042-162053. PMID: 36754333
Handzlik, M.K.; Gengatharan, J.M.; Frizzi, K.E.; et al. 2023. Insulin-regulated serine and lipid metabolism drive peripheral neuropathy. Nature, 614(7946), 118-124. PMID: 36697822
Shen, J.L.; Doherty, J.; Allen, E.; et al. 2022. Atg6 promotes organismal health by suppression of cell stress and inflammation. Cell Death Differ, 29(11), 2275-2287. PMID: 35523956
Fuenzalida, K.; Leal-Witt, M.J.; Guerrero, P.; et al. 2021. NTBC treatment monitoring in Chilean patients with tyrosinemia type 1 and its association with biochemical parameters and liver biomarkers. J Clin Med, 10(24), 5832-5845. PMID: 34945128
Lim, E.W.; Handzlik, M.K.; Trefts, E.; et al. 2021. Progressive alterations in amino acid and lipid metabolism correlate with peripheral neuropathy in PolgD257A mice. Sci Adv, 7(42), eabj4077. PMID: 34652935
Gebert, N.; Rahman, S.; Lewis C.A.; et al. 2021. Identifying cell-type-specific metabolic signatures using transcriptome and proteome analyses. Curr Protoc, 1(9), e245. PMID: 34516047
Gauthier-Coles, G.; Vennitti, J.; Zhang, Z.; et al. 2021. Quantitative modelling of amino acid transport and homeostasis in mammalian cells. Nat Commun, 12(1), 5282-5300. PMID: 34489418
Liu, Y.; Zhao, Y.; Shukha, Y.; et al. 2021. Dysregulated oxalate metabolism is a driver and therapeutic target in atherosclerosis. Cell Rep, 36(4), 109420-109457. PMID: 34320345
Gao, J.; Chung, T-S. 2021. Membranes made from nonsolvent-thermally induced phase separation (N-TIPS) for decellularization of blood in dry plasma spot (DPS) applications. Chem Eng Sci, 229. doi.org/10.1016/j.ces.2020.116010.
Kimhofer, T.; Lodge, S.; Whiley, L.; et al. 2020. Integrative modeling of quantitative plasma lipoprotein, metabolic, and amino acid data reveals a multiorgan pathological signature of SARS-CoV-2 infection. J Proteome Res, 19(11), 4442-4454. PMID: 32806897
van Gastel, N.; Spinelli, J.B.; Sharda, A.; et al. 2020. Induction of a timed metabolic collapse to overcome cancer chemoresistance. Cell Metab, 32(3), 391-403. PMID: 32763164
Røst, L.M.; Brekke Thorfinnsdottir, L.; Kumar, K.; et al. 2020. Absolute quantification of the central carbon metabolome in eight commonly applied prokaryotic and eukaryotic model systems. Metabolites, 10(2), 74. PMID: 32093075
Alcock, R.D.; Shaw, G.C.; Tee, N.; et al. 2019. Plasma amino acid concentrations after the ingestion of dairy and collagen proteins, in healthy active males. Front Nutr, 6, 163. PMID: 31681789
Sullivan, M.R; Danai, L.V.; Lewis, C.A.; et al. 2019. Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability. Elife, 8, e44235. PMID: 30990168
Lu, Y.; Wang, Y.; Liang, X.; et al. 2019. Serum amino acids in association with prevalent and incident type 2 diabetes in a Chinese population. Metabolites, 9(1). PMID: 30646552
Haines, N.R.; Manoharan, N.; Olson, J.L.; et al. 2018. Metabolomics analysis of human vitreous in diabetic retinopathy and rhegmatogenous retinal detachment. J Proteome Res, 17(7), 2421-2427. PMID: 29877085
Chen, W.W.; Freinkman, E.; Sabatini, D.M. 2017. Rapid immunopurification of mitochondria for metabolite profiling and absolute quantification of matrix metabolites. Nat Protoc, 12(10), 2215-2231. PMID: 29532801
Pretorius, C.J.; McWhinney, B.C.; Sipinkoski, B.; et al. 2018. Rapid amino acid quantitation with pre-column derivatization; ultra-performance reverse phase liquid chromatography and single quadrupole mass spectrometry. Clin Chim Acta, 478, 132-139. PMID: 29274329
Mehta, K.Y.; Wu, H.J.; Menon, S.S.; et al. 2017. Metabolomic biomarkers of pancreatic cancer: a meta-analysis study. Oncotarget, 8(40), 68899-68915. PMID: 28978166
Havelund, J.F.; Andersen, A.D.; Binzer, M.; et al. 2017. Changes in kynurenine pathway metabolism in Parkinson patients with L-DOPA-induced dyskinesia. J Neurochem, 142(5), 756-766. PMID: 28628213
Nemkov, T.; Hansen, K.C.; D'Alessandro, A. 2017. A three-minute method for high-throughput quantitative metabolomics and quantitative tracing experiments of central carbon and nitrogen pathways. Rapid Commun Mass Spectrom, 31(8), 663-673. PMID: 28195377
D'Alessandro, A.; Nemkov, T..; Yoshida, T.; et al. 2017. Citrate metabolism in red blood cells stored in additive solution-3. Transfusion, 57(2), 325-336. PMID: 27813142
Mayers, J.R.; Torrence, M.E.; Danai, L.V.; et al. 2016. Tissue of origin dictates branched-chain amino acid metabolism in mutant Kras-driven cancers. Science, 353(6304), 1161-1165. PMID: 27609895
Chen, W.W.; Freinkman, E.; Wang, T.; et al. 2016. Absolute quantification of matrix metabolites reveals the dynamics of mitochondrial metabolism. Cell, 166(5), 1324-1337. PMID: 27565352