13c6 15n2 lysine

Manufactured by Merck Group

Sourced in France

13C6,15N2-lysine is a stable isotope-labeled amino acid used as a research tool in various analytical techniques. It provides a means to track and quantify proteins and their modifications in biological samples. The compound consists of a lysine molecule with six carbon-13 atoms and two nitrogen-15 atoms, allowing for accurate mass spectrometry-based analysis.

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Lab products found in correlation

13c6 15n4 arginine, by Merck Group (2 mentions) Mca rppgfsafk dnp oh, by R&D Systems (1 mentions) Flexmap 3d instrument, by Merck Group (1 mentions) Chymotrypsin, by Merck Group (1 mentions) Peptidase from porcine intestinal mucosa, by Merck Group (1 mentions) Trypsin, by Merck Group (1 mentions) Ptrchisa vector, by Thermo Fisher Scientific (1 mentions) Low glucose dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Geneticin, by Thermo Fisher Scientific (1 mentions) Formic acid, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Dialyzed fbs, by Gemini Bio (1 mentions) Bca protein quantification kit, by Thermo Fisher Scientific (1 mentions) 13c6 arginine, by Cambridge Isotopes (1 mentions)

Close spelling variants of this product

[13C6,15N2]L-lysine-HCl [13C6,15N2]-L-lysine 13C6 lysine

6 protocols using 13c6 15n2 lysine

In Vitro Selection of IDE-Targeting Macrocyclic Inhibitors

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The in vitro selection of the DNA-templated library^{8 (link)} used 20 µg His₆-tagged mouse IDE immobilized on cobalt magnetic beads (Invitrogen). IDE inhibition was assayed using the fluorogenic peptide Mca-RPPGFSAFK(Dnp)-OH (R&D), confirmed using an anti-insulin antibody time-resolved FRET assay (Cysbio), and a LCMS assay for CGRP cleavage fragments in plasma^{9 (link)}. Macrocyclic inhibitors were synthesized by Fmoc-based solid-phase synthesis and purified by HPLC. LCMS quantitation of 6bK in biological samples was performed using 6bK synthesized with ¹³C₆,¹⁵N₂ lysine (Sigma-Aldrich).
Wild-type lean and DIO C57BL/6J age-matched male mice (Jackson Laboratories) were used at 14–16, and 24–26 weeks respectively (> 20 weeks of high-fat diet). Gcgr^−/− and Ide^−/− mice were fully backcrossed to the C57BL/6J line, bred from heterozygous mice, and used between 11 and 21 weeks. Animals were fasted overnight 14 h for all experiments, except for the insulin tolerance test, which required 5 h of fasting during the morning. Blood glucose was measured from tail nicks using AccuCheck (Aviva) meters. Trunk blood was obtained for plasma hormone measurements using the Multiplexed Mouse Metabolic Hormone panel (Milliplex, EMD Millipore) on a Luminex FlexMap 3D instrument.

Maianti J.P., McFedries A., Foda Z.H., Kleiner R.E., Du X.Q., Leissring M.A., Tang W.J., Charron M.J., Seeliger M.A., Saghatelian A, & Liu D.R. (2014). Anti-diabetic activity of insulin-degrading enzyme inhibitors mediated by multiple hormones. Nature, 511(7507), 94-98.

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Rapeseed Meal Preparation and Analysis

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Rapeseed meals were prepared from 00-rapeseed (Brassica napus) at the pilot plant of CREOL/OLEAD (Pessac, France). Trypsin (type IX-S, 13,000–20,000 BAEE units/mg protein, EC 232-650-8), chymoTrypsin (type II, ≥40 units/mg protein, EC 232-671-2) and peptidase from porcine intestinal mucosa (50–100 units/g solid, EC 232-875-1) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The furosine, lysinoalanine and N^ε-carboxymethyl-lysine standards were obtained from PolyPeptide Laboratories (Strasbourg, France), whereas the rest of the standards (¹³C₆,¹⁵N₂-lysine, lysine, lanthionine) were obtained from Sigma-Aldrich (Steinheim, Germany).

Salazar-Villanea S., Bruininx E.M., Gruppen H., Carré P., Quinsac A, & van der Poel A.F. (2017). Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins. Journal of the American Oil Chemists' Society, 94(4), 619-630.

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Stable Isotope Labeling of Ras Proteins

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¹³C₆-¹⁵N₂-lysine (+8 Da) and ¹³C₆-¹⁵-N₄-arginine (+10 Da) were obtained from Sigma. His-tagged, wild-type KRAS4B and NRAS plasmids were kindly provided by Ignacio Rubio (Institute of Molecular Cell Biology, University of Jena). Wild-type, full-length human HRAS and KRAS4A sequences were sub-cloned into the pTrcHis A vector (Invitrogen) from plasmids described in [23 (link), 24 (link)] and sequence verified.

Mageean C.J., Griffiths J.R., Smith D.L., Clague M.J, & Prior I.A. (2015). Absolute Quantification of Endogenous Ras Isoform Abundance. PLoS ONE, 10(11), e0142674.

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Animal Experiment Protocols and Standards

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The Central Committee of Animal Experiments (The Netherlands) approved the use of experimental animals under the authorization number AVD260002015139. All animals were euthanized under sodium pentobarbital anesthesia, and all efforts were made to minimize animal suffering. The furosine, LAL and CML standards were obtained from Polypeptides (Strasbourg, France), whilst the rest of the standards (¹³C₆,¹⁵N₂-lysine, lysine, LAN) were obtained from Sigma-Aldrich (Steinheim, Germany).

Salazar-Villanea S., Butré C.I., Wierenga P.A., Bruininx E.M., Gruppen H., Hendriks W.H, & van der Poel A.F. (2018). Apparent ileal digestibility of Maillard reaction products in growing pigs. PLoS ONE, 13(7), e0199499.

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SILAC-Based Membrane Protein Quantification

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Heavy amino acids, ¹³C₆¹⁵N₂-lysine and ¹³C₆¹⁵N₄-arginine, were obtained from Sigma-Aldrich (St. Louis, MO). Low-glucose Dulbecco's modified Eagle's medium (DMEM), phosphate-buffered saline (PBS), penicillin/streptomycin, and geneticin were purchased from Invitrogen, Carlsbad, CA. Fetal bovine serum (FBS) and dialyzed FBS were procured from Gemini Bio-Products, Calabasas, CA. SILAC DMEM (without L-lysine and L-arginine) was purchased from Thermo Fisher Scientific, MA, USA. The ProteoExtract Native Membrane Protein Extraction Kit was purchased from Calbiochem (Temecula, CA). The protein quantification BCA kit and the in-solution digestion kit were from Pierce Biotechnology (Rockford, IL). Pure synthetic unlabeled peptides, NVTGFFQSFK, NVTGFFQSLK, VLAVTDSPAR, and NTTGALTTR, with purity determined by quantitative amino acid analysis, were obtained from New England Peptides (Boston, MA). The SIL internal standards, NVTGFFQSF[¹³C₆¹⁵N₂]K, NVTGFFQSL[¹³C₆¹⁵N₂]K, VLAVTDSPA[¹³C₆¹⁵N₄]R, and NTTGALTT[¹³C₆¹⁵N₄]R, were from Thermo Fisher Scientific (Rockford, IL). HPLC-grade acetonitrile and other solvents were purchased from Fischer Scientific (Fair Lawn, NJ) and formic acid was purchased from Sigma-Aldrich (St. Louis, MO).

Prasad B, & Unadkat J.D. (2014). Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MS/MS Quantification of Hepatic Drug Transporters. International Journal of Proteomics, 2014, 451510.

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Antibody and Isotope Labeling Protocol

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Antibodies used in the study are listed in Supplemental Table S2. The ¹³C₆ arginine was from the Cambridge Isotope Laboratories, Tewksbury, MA; ¹³C₆-¹⁵N₂ lysine was from Sigma-Aldrich, St. Louis, MO.

Mueller S., Wahlander A., Selevsek N., Otto C., Ngwa E.M., Poljak K., Frey A.D., Aebi M, & Gauss R. (2015). Protein degradation corrects for imbalanced subunit stoichiometry in OST complex assembly. Molecular Biology of the Cell, 26(14), 2596-2608.

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