13c615n2 lysine

Manufactured by Thermo Fisher Scientific

[13C615N2] lysine is a stable isotope-labeled amino acid. It is used as a tracer for studying protein synthesis and turnover in biological systems.

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Lab products found in correlation

13c615n4 arginine, by Thermo Fisher Scientific (2 mentions) Iodoacetamide, by GE Healthcare (1 mentions) Arg c, by Promega (1 mentions) 660 nm protein assay kit, by Thermo Fisher Scientific (1 mentions) High capacity streptavidin agarose beads, by Thermo Fisher Scientific (1 mentions) Ms grade trypsin, by Promega (1 mentions) Horseradish peroxidase hrp conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Hrp conjugated species specific secondary antibodies, by Cell Signaling Technology (1 mentions) Rpmi 1640 medium for silac, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Supersignal chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Dmem media, by Thermo Fisher Scientific (1 mentions) Chloroform, by Merck Group (1 mentions) Methanol, by Merck Group (1 mentions) Ammonium formate, by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Ammonium bicarbonate, by Merck Group (1 mentions) Asp n, by Promega (1 mentions)

3 protocols using 13c615n2 lysine

Quantification of Tau Protein by SRM-MS

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The SID-SRM-MS assays of selected proteins were developed as described previously (85 (link)). For protein tau, two or three peptides were initially selected, and then the sensitivity and selectivity of these were experimentally evaluated as described previously (85 (link)). The peptide with the best sensitivity and selectivity was selected as the surrogate for that protein. For each peptide, 3–5 SRM transitions were monitored (Table S2). The signature peptides of various ubiquitination linkages were chemically synthesized, incorporating isotopically labeled [¹³C₆¹⁵N₄] arginine or [¹³C₆¹⁵N₂] lysine to a 99% isotopic enrichment (Thermo Scientific). The amount of stable isotope-labeled standard (SIS) peptides was determined by amino acid analysis. After trypsin digestion, the tau peptides were then reconstituted in 30 μl of 5% formic acid-0.01% TFA, and an aliquot of SIS peptides was added to each tryptic digest. These samples were desalted with a ZipTip C18 cartridge and analyzed by LC-SRM-MS as described above.

Puangmalai N., Sengupta U., Bhatt N., Gaikwad S., Montalbano M., Bhuyan A., Garcia S., McAllen S., Sonawane M., Jerez C., Zhao Y, & Kayed R. (2022). Lysine 63-linked ubiquitination of tau oligomers contributes to the pathogenesis of Alzheimer’s disease. The Journal of Biological Chemistry, 298(4), 101766.

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Quantitative Proteomic Analysis of Cell Signaling

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Iodoacetamide was purchased from GE Healthcare. RPMI-1640 and DMEM media were from Gibco. Microcon YM-30 spin filters were from Millipore. MS-grade Trypsin, Asp-N, and Arg-C were from Promega. Tris, SDS, methanol, chloroform, 2,2′-dithiodipyridine (DTDP), dime-thylformamide (DMF), hydroxylamine, sodium chloride, ammonium bicarbonate, and ammonium formate were from Sigma-Aldrich. Arginine (Arg0), lysine (Lys0), ¹³C₆¹⁵N₄-arginine (Arg10), ¹³C₆¹⁵N₂-lysine (Lys8), dialyzed fetal bovine serum, RPMI 1640 medium for SILAC, 660 nm Protein Assay Kit, TCEP, NEM, high-capacity streptavidin agarose beads, SuperSignal chemiluminescent substrate, trap columns, EASY-Spray analytical columns, and Horseradish peroxidase (HRP)-conjugated streptavidin were from Thermo Fisher Scientific. Primary antibodies were from Cell Signaling Technology, Novus Biologicals, Santa Cruz Biotechnology, Sigma-Aldrich, and R&D Systems. HRP-conjugated species-specific secondary antibodies were from Cell Signaling Technology and Jackson ImmunoResearch Laboratory.

Zhou B., Wang Y., Yan Y., Mariscal J., Di Vizio D., Freeman M.R, & Yang W. (2019). Low-Background Acyl-Biotinyl Exchange Largely Eliminates the Coisolation of Non-S-Acylated Proteins and Enables Deep S-Acylproteomic Analysis. Analytical chemistry, 91(15), 9858-9866.

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Quantification of Retinaldehyde Metabolism Enzymes

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Retinaldehyde (RAL), all‐trans‐retinoic acid (atRA) and nicotinamide adenine dinucleotide (NAD⁺) were purchased from Millipore Sigma (Burlington, MA) and atRA‐d₅ from Cambridge Isotope Laboratories (Tewksbury, MA). All mass spectrometry grade solvents were purchased from Thermo Fisher Scientific (Waltham, MA). Stable isotope labeled (SIL) peptides labeled with [¹³C₆¹⁵N₂] arginine and [¹³C₆¹⁵N₂] lysine were from Thermo Fisher Scientific (Waltham, MA) and used as internal standards (IS) for AOX, aldehyde dehydrogenase 1 family member A1 (ALDH1A1) and family member A2 (ALDH1A2) protein quantification.²¹, ²² Human recombinant ALDH1A1, ALDH1A2, and AOX were expressed and purified as described previously and used as quantification standards.²¹

Rubinow K.B., Zhong G., Czuba L.C., Chen J.Y., Williams E., Parr Z., Khandelwal S., Kim D., LaFrance J, & Isoherranen N. (2022). Evidence of depot‐specific regulation of all‐trans‐retinoic acid biosynthesis in human adipose tissue. Clinical and Translational Science, 15(6), 1460-1471.

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