L asparagine

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom

L-asparagine is a non-essential amino acid used as a laboratory reagent. It is a white crystalline solid at room temperature and has the chemical formula C4H8N2O3.

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Lab products found in correlation

103 protocols using l asparagine

Amino Acid Standard Preparation Protocol

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Sodium acetate trihydrate (pro analysis), acetic acid (glacial), acetonitrile (HPLC grade), and methanol (HPLC grade) were purchased from Merck KGaA (Darmstadt, Germany). Disodium hydrogen orthophosphate heptahydrate (98.0–102.0%), ethylenediamine tetraacetic acid (EDTA) (>99%), and orthophosphoric acid (>85%) were purchased from VWR Chemicals BDH (Radnor, PA, USA). Triethylamine (>99%), DL-norleucine (98%), L-asparagine (>98%), L-glutamine (>99%), L-tryptophan (>98%), DL-dithiothreitol (>985), and phenylisothiocyanate (>99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
The following reference materials were purchased from Sigma-Aldrich: Amino acid standard solution AAS18 (2.5 μmol mL⁻¹ of 18 proteinogenic amino acids in 0.1 N HCl), Amino acid standard solution A6407 (2.5 μmol mL⁻¹ of 26 physiological amino acids in 0.1 N HCl), Amino acid standard solution A6282 (2.5 μmol mL⁻¹ of 14 physiological, basic amino acids in 0.1 N HCl).
A proteinogenic amino acid standard consisting of Amino acid standard solution AAS18 and 0.05 μmol mL⁻¹ of L-asparagine, L-glutamine and L-tryptophan was prepared. A composite physiological amino acid standard was prepared from equal volumes of amino acid standard solutions A6407 and A6282.

Milan A.M., Samuelsson L.M., Shrestha A., Sharma P., Day L, & Cameron-Smith D. (2020). Circulating Branched Chain Amino Acid Concentrations Are Higher in Dairy-Avoiding Females Following an Equal Volume of Sheep Milk Relative to Cow Milk: A Randomized Controlled Trial. Frontiers in Nutrition, 7, 553674.

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Development of a μPAD Sensor

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All reagents used were of analytical reagent grade unless stated otherwise and were obtained from Sigma-Aldrich (Sigma-Aldrich Química S.A., Madrid, Spain). All aqueous solutions were made using reverse osmosis-type quality water, Milli-RO 12 plus Milli-Q station from Millipore, with a conductivity of 18.2 MΩ·cm (Millipore, Bedford, MA, USA). 3,3′,5,5′-tetramethylbenzidine (TMB) (CAS No, 54827-17-7); acetic acid (CAS No, 64-19-7); sodium acetate (CAS No, 127-09-3); ethanol (CAS No, 64-17-5); L-glutathione (GSH) (CAS No, 70-18-8); L-cysteine (Cys) (CAS No, 7048-04-6); L-glutamine (Gln) (CAS No, 56-85-9); L-arginine (Arg) (CAS No, 74-79-3); L-asparagine (Asn) (CAS No, 70-47-3); DL-homocysteine (Hcys) (CAS No, 454-29-5); L-glutamic acid (Glu) (CAS No, 56-86-0); D-mannose (Man) (CAS No, 3458-28-4); galactose (Gal) (CAS No, 137868-52-1); D-lysine (Lys) (CAS No, 923-27-3); L-serine (Ser) (CAS No, 56-45-1); L-histidine (His) (CAS No, 71-00-1); Bovine serum albumin (BSA) (CAS No, 9048-46-8); and Silver nitrate (AgNO₃) (CAS No, 7761-88-8) were acquired from Merck (Darmstadt, Germany). Filter paper (ref. Whatman 1, qualitative circles 150 mm) from Sigma-Aldrich and 54 × 86 mm transparent sheets with a quality strength protection of 125 microns (Fellowes^® pouches, Itaca, NY, USA) were used to prepare and laminate the μPAD.

Ortiz-Gómez I., Rivadeneyra A., Salmerón J.F., de Orbe-Payá I., Morales D.P., Capitán-Vallvey L.F, & Salinas-Castillo A. (2023). Near-Field Communication Tag for Colorimetric Glutathione Determination with a Paper-Based Microfluidic Device. Biosensors, 13(2), 267.

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Establishment of Melanoma Cell Lines

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P511 is an azaguanine-resistant variant of P815 and P1.204 is a P815AB-negative variant, carrying a deletion of gene P1A (official gene name Trap1a).37 (link) L1210.P1A.B7-1 cells were obtained by transfection of L1210.P1A leukemia cells with the murine B7-1 cDNA cloned into plasmid pEFBOS.38 (link) All cells were maintained at 37°C with 8% CO₂. Unless otherwise specified, all culture media contained 10% FBS supplemented with L-Arginine (0.55 mM, Merck), L-asparagine (0.24 mM, Merck), Glutamine (1.5 mM, Merck), beta-mercaptoethanol (50 µM, Sigma), 50 U mL−1 penicillin and 50 mg mL−1 streptomycin (Life Technologies). Melanoma cells T429.11 were derived from an Amela TiRP tumor.33 (link) Cell lines were routinely tested for mycoplasma contamination. Primary melanoma cell line Xni-5 was established from a Mela TiRP tumor, using γ-irradiated mouse keratinocytes (XB-2 line obtained from ATCC) as feeder cells, and was grown in culture medium containing 10% FBS supplemented with 12-O-Tetradecanoylphorbol 13-acetate (10 ng/mL, WAKO) and cholera toxin (10 nM, Sigma-Aldrich, St Louis, Missouri, USA). Freshly irradiated keratinocytes were added in the co-culture every week up to passage 3 (3 weeks after initial plating).

Canè S., Van Snick J., Uyttenhove C., Pilotte L, & Van den Eynde B.J. (2021). TGFβ1 neutralization displays therapeutic efficacy through both an immunomodulatory and a non-immune tumor-intrinsic mechanism. Journal for Immunotherapy of Cancer, 9(2), e001798.

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Enzymatic Assay for Oxidative Enzymes

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Syringaldazine (4-hydroxy-3,5-dimethoxybenzaldehyde azine), ferulic acid (4-hydroxy-3-methoxycinnamic acid), and veratric acid (3,4-dimethoxybenzoic acid) were supplied by Sigma-Aldrich (St. Louis, MO, USA), while l-asparagine was purchased from Merck (Darmstadt, Germany). All other products used were of reagent or analytical grade and purchased locally.

Janusz G., Czuryło A., Frąc M., Rola B., Sulej J., Pawlik A., Siwulski M, & Rogalski J. (2014). Laccase production and metabolic diversity among Flammulina velutipes strains. World Journal of Microbiology & Biotechnology, 31(1), 121-133.

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Cell Maintenance and Hypoxic Culture

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L1210.P1A.B7-1, P511 and P1.204 cell lines were already described.30 (link) MC38 cells were a gift from P. Berraondo (Navarra); LL2-Thy1.1-OVA (LLC-OVA) cells, expressing a cytoplasmic form of ovalbumin, were a gift from D. Fearon (Cambridge); OVA-transfected EL-4 cells (EG7) were a gift from J. Van Ginderachter (Brussels); B16F1 cells were obtained from ATCC (CRL-6323). T429.11 clone was derived from an inducible TiRP melanoma as already described.30 (link)
All cells were maintained at 37°C and 8% CO₂. All culture media contained 10% fetal bovin serum, L-arginine (0.55 mM, Merck), L-asparagine (0.24 mM, Merck), glutamine (1.5 mM, Merck), 50 U/mL penicillin and 50 mg/mL streptomycin (Life Technologies) (complete medium). Cell lines were routinely tested for mycoplasma contamination. For experiments in hypoxia, a hypoxystation (Whitley H35) at 37°C, 8% CO₂ and 1% O₂ was used.

Finisguerra V., Dvorakova T., Formenti M., Van Meerbeeck P., Mignion L., Gallez B, & Van den Eynde B.J. (2023). Metformin improves cancer immunotherapy by directly rescuing tumor-infiltrating CD8 T lymphocytes from hypoxia-induced immunosuppression. Journal for Immunotherapy of Cancer, 11(5), e005719.

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Purification of L-Asparaginase from E.coli

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L-asparagine, L-glutamine, Trichloro acetic acid and Tris were purchased from Merck (Darmstadt, Germany). DEAE-Sepharose was provided by Pharmacia (Uppsala, Sweden). L-asparaginase derived from E.coli was a gift from cancer research institute, Iran. All other chemicals were from Sigma (St. Louis, MO, USA) and were of analytical grade.

Rahimzadeh M., Poodat M., Javadpour S., Qeshmi F.I, & Shamsipour F. (2016). Purification, Characterization and Comparison between Two New L-asparaginases from Bacillus PG03 and Bacillus PG04. The Open Biochemistry Journal, 10, 35-45.

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Characterization of T429 Melanoma Clones

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T429.11, T429.6 and T429.18 clones were derived from an induced Amela TiRP tumor referred to as T429^{4 (link)}. They were cloned from the T429 induced melanoma primary tumor line. The expression of the transgene^{2 (link)} was evaluated in all three T429 clones and T-cell recognition was tested for each of them in vitro using TCRP1A CD8⁺ T cells. P511 is an azaguanine-resistant variant of P815^{48 (link), 49 (link)}. P1204 is a P815AB-negative variant carrying a deletion of gene Trap1a^{48 (link), 49 (link)}. L1210.P1A.B7-1 cells were obtained by transfection of L1210.P1A cells with the murine B7-1 cDNA cloned into plasmid pEFBOS^{50 (link)}. All cells were maintained at 37 °C with 8% CO₂. Unless otherwise specified, all culture media are IMDM (GIBCO) containing 10% fetal bovine serum supplemented with L-arginine (0.55 mM, Merck), L-asparagine (0.24 mM, Merck), glutamine (1.5 mM, Merck), beta-mercaptoethanol (50 µM, Sigma), 50 U ml⁻¹ penicillin and 50 mg ml⁻¹ streptomycin (Life Technologies) (complete medium). Cell lines were tested for mycoplasma contamination.

Zhu J., Powis de Tenbossche C.G., Cané S., Colau D., van Baren N., Lurquin C., Schmitt-Verhulst A.M., Liljeström P., Uyttenhove C, & Van den Eynde B.J. (2017). Resistance to cancer immunotherapy mediated by apoptosis of tumor-infiltrating lymphocytes. Nature Communications, 8, 1404.

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Biochemical Composition Analysis

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Sucrose (the sugar), the bitter caffeine, the amino acids L-Aspartic acid, L-Glutamic acid, L-Glutamine, L-Lysine, L-Asparagine, L-Arginine, L-Methionine, L-Glycine, L-Threonine, L-Valine, L-Proline, L-Leucine, L-Phenylalanine, L-Histidine, L-Isoleucine, L-Serine, L-Glutamic acid monosodium salt (MSG), the ribonucleotides Inosine 5′-monophosphate (IMP) and Guanosine 5′-monophosphate (GMP) were purchased from Sigma Aldrich (Milwaukee,USA). All the chemicals are of analytical grade (>99.5%).

Lim S., Jung J., Yunusbaev U., Ilyasov R, & Kwon H.W. (2019). Characterization and its implication of a novel taste receptor detecting nutrients in the honey bee, Apis mellifera. Scientific Reports, 9, 11620.

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Analytical Standards for Tea Bioactives

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Catechin (C, ≥98%), galloCatechin (GC, ≥98%), galloCatechin-3-gallate (GCG, ≥98%), epiCatechin (EC, ≥98%), epiCatechin-3-gallate (ECG, ≥98%), epigalloCatechin (EGC, ≥98%), epigalloCatechin-3-gallate (EGCG, ≥98%), caffeine (CAF, ≥98%), theaflavins (TF, ≥95%), theaflavins-3-gallate (TF3G, ≥98%), theaflavin-3′-gallate (TF3′G, ≥98%), theaflavine-3,3′-digallate (TFDG, ≥98%), quercetin-3-O-galactoside (≥98%), myricetin-3-O-galactoside (≥98%), kaempferol-3-O-rutinoside (≥98%), vitexin (≥98%), astragaloside (≥98%), quercetin (≥98%), and kaempferol (≥98%) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China). l-alanine, l-arginine, l-aspartic acid, l-cystine, l-glutamic acid, glycine, l-histidine, l-isoleucine, l-leucine, l-lysine, l-methionine, l-phenylalanine, l-proline, l-serine, l-threonine, l-tyrosine, l-valine, l-asparagine, l-glutamine, theanine, γ-aminobutyric acid, and l-tryptophan were purchased from Sigma (Sigma-Aldrich Co., St. Louis, MO, USA). Anthrone reagent and ninhydrin/formic acid reducing agent were purchased from BeiJing DingGuochangSheng Biotech. Co., Ltd. (Bejing, China). Ethyl caprate (≥99%) was purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China), and n-alkane mixed standard C7-C40 was purchased from o2si (o2si smart solutions—an LGC Standards Company, Charleston, SC, USA).

Lin J., Wang Y., Chen L., Yang Y., Tu Z, & Ye Y. (2022). Effect of the Presence of Stem on Quality of Oolong Tea. Foods, 11(21), 3439.

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Amino Acid Analysis by UHPLC

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Sulfuric acid, sodium hydroxide, acetonitrile (Ultra high performance liquid chromatography (UHPLC) grade), ethylenediaminetetraacetic acid (EDTA), sodium borate, and hydrochloric acid were obtained from Panreac (Castelar del Vallés, Barcelona, Spain). Sodium acetate (anhydrous), trimethylamine (TEA), phosphoric acid, ammonium molybdate, and methyl cellulose were purchased from Sigma-Aldrich (Darmstadt, Germany). Standard of amino acids (L-arginine (Arg), L-alanine (Ala), L-asparagine (Asn), L-aspartic acid (Asp), glycine (Gly), L-glutamic acid (Glu), L-glutamine (Gln), L-histidine (His), L-iso Leucine (Ileu), Leucine (Leu), L-Lysine (Lys), L-norvaline (Nor), L-phenylalanine (Phe), L-proline (Pro), L-serine (Ser), L-threonine (Thr), L-tryptophan (Trp), L-tyrosine (Tyr), and L-valine (Val)) were purchased from Sigma-Aldrich (Steinheim, Germany). All the other chemicals and reagents used were of analytical grade.

Carbas B., Machado N., Oppolzer D., Ferreira L., Queiroz M., Brites C., Rosa E.A, & Barros A.I. (2020). Nutrients, Antinutrients, Phenolic Composition, and Antioxidant Activity of Common Bean Cultivars and their Potential for Food Applications. Antioxidants, 9(2), 186.

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