Taurine–glycine resolving gel composition: 75 mM tris (pH = 10) (Euromedex, Strasbourg, France), 200 mM Taurine, 125 mM glycine (Euromedex, Strasbourg, France), 23 mM HCl, 12% acryl/bisacryl 37.5:1 (Biosolve, Dieuze, France). Taurine–asparagine resolving gel composition: 75 mM tris–HCl (pH = 8.8, ), 100 mM Taurine, 100 mM asparagine (Sigma-Aldrich, St. Louis, MO, USA), 12% acryl/bisacryl 37.5:1. glycine–asparagine resolving gel composition: 75 mM tris–HCl (pH = 8.8), 125 mM glycine, 150 mM asparagine, 23 mM HCl, 12% acryl/bisacryl 37.5:1. SDS-PAGE (tris–glycine) resolving gels composition: 380 mM tris–HCl pH = 8.8, 0.1% SDS (Sigma-Aldrich), 12% acryl/bisacryl 37.5:1. Gel polymerization was induced by the addition of APS (0.6 mg/mL, Sigma-Aldrich) and Temed (1 µL/mL, Sigma-Aldrich). Stacking mini-gels formulation: 125 mM tris–HCl pH = 6.7 and 5% acryl/bisacryl 37.5:1. In the case of SDS-PAGE, 0.1% SDS was added. Migration buffer for all the electrophoretic separations was glycine 14.4 g/L, tris 3 g/L, SDS 1 g/L, pH = 8.8. Electrophoretic conditions are set at 25 mA per mini-gel.
Asparagine
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom
Asparagine is a laboratory-grade amino acid used in various biochemical and cell culture applications. It serves as a nutrient and a precursor for the synthesis of other amino acids and biomolecules. Asparagine is a commonly used component in cell culture media, supporting the growth and maintenance of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Glutamine, by Merck Group (29 mentions)
Tryptophan, by Merck Group (23 mentions)
Aspartic acid, by Merck Group (20 mentions)
Serine, by Merck Group (19 mentions)
Phenylalanine, by Merck Group (19 mentions)
Alanine, by Merck Group (19 mentions)
Glutamic acid, by Merck Group (18 mentions)
Arginine, by Merck Group (17 mentions)
Histidine, by Merck Group (17 mentions)
Valine, by Merck Group (17 mentions)
Methionine, by Merck Group (16 mentions)
Leucine, by Merck Group (16 mentions)
Glycine, by Merck Group (15 mentions)
Threonine, by Merck Group (15 mentions)
Proline, by Merck Group (15 mentions)
Isoleucine, by Merck Group (14 mentions)
Cysteine, by Merck Group (14 mentions)
Lysine, by Merck Group (14 mentions)
Tyrosine, by Merck Group (12 mentions)
Methanol, by Merck Group (9 mentions)
78 protocols using asparagine
1
Optimized Gel Compositions for Protein Separation
2
Optimized Bacterial Growth Media Formulation
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The reagents used were Ampicillin, Al2(SO4)3·16H2O, thiamine·HCl, ZnSO4·7H2O, IPTG, Tris, hydroxylamine hydrochloride, Asparagine, FeCl3, TCA and L-aspartic acid β-hydroxamate were Sigma-Aldrich™ (Steinheim, NW, Germany). The reagents KH2PO4, citric acid, MgSO4·7H2O, K2HPO4, ZnCl2, CaCl2·2H2O, CuSO4·5H2O, FeSO4·7H2O, NH4OH, HCl and lactose were Vetec Quimica Fina™ (Duque de Caxias, RJ, Brazil). The reagents glucose, (NH4)2HPO4, MnCl2·4H2O, Zn (CH3COO)2·H2O, CoCl2·6H2O, CuCl2·2H2O, EDTA·Na2, MnSO4·H2O, NiSO4·6H2O, Acetate and H2SO4 were Dinâmica Quimica™ (Indaíatuba, SP, Brazil). The reagents tryptone was Acumedia™, yeast extract was Kasvi™ (São José dos Pinhais, PR, Brazil), sodium chloride was Cromoline™ (Diadema, SP, Brazil), Glycerol was PlusOne™, Iron (III) Citrate was Êxodo científica™ (Sumaré, SP, Brazil), H3BO3 was Quimex™ (Uberaba, MG, Brazil), Na2MoO4·2H2O was Merck™ (Darmstadt, Germany) and NaH2PO4·H2O was labsynth™ (Diadema, SP, Brazil).
3
Deriving Immortalized Pancreatic Fibroblasts
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pMAFs were derived from mCherry-GFP-LC3 homozygous mice (kindly provided by Ian Ganley, The University of Dundee). After aseptic dissection, the pancreas was finely minced and digested in a “liberase blendzyme” (Roche, 05401020001) for 60 min at 37°C. Tissue fragments were then plated onto 10-cm culture dishes and allowed to attach. Fibroblasts were seen to “crawl” out of the tissue for up to 10 d. These fibroblasts were harvested and sub-cultured in RPMI 1640 medium (GlutaMAX supplemented, Life Technologies, 72,400,021) with 10% FCS, 50 µM of 2-mercaptoethanol (Sigma Aldrich, M6250) and 50 µM of asparagine (Sigma Aldrich, A4159). The fibroblasts divide rapidly for a few divisions, then slow down and cell numbers remain constant or decline slightly for up to 6–8 weeks. Following on from this “crisis” the cells once again return to fast replication at which point they are considered immortal and suitable for use in ongoing studies.
4
Culturing Mouse and Human Lymphoma Cells
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Eμ-Myc mouse lymphoma cells were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Gibco), 50 μM β-mercaptoethanol (Sigma-Aldrich), 100 μM asparagine (Sigma-Aldrich), 100 U/mL penicillin, and 100 mg/mL streptomycin (Gibco) at 37°C and 10% CO2. OP9 cells were cultured in αMEM (Gibco) supplemented with 20% heat-inactivated FBS, 1 mM glutamine (Gibco), 10 mM Hepes (Gibco), 1 mM sodium pyruvate (Gibco), 50 μM β-mercaptoethanol, 100 U/mL penicillin, and 100 μg/mL streptomycin. Human cell lines were cultured in a humidified incubator at 37°C and 5% CO2. Virus-producing 293T cells were maintained in DMEM supplemented with 10% FBS. Approximately 6 h prior to transfection, 293T cells were cultured in DME glutamax (Gibco) supplemented with 10% FBS and 25 mM Hepes. Rael-BL, Ramos-BL, Sav-BL, and BL-31 human Burkitt lymphoma-derived cell lines were cultured in RPMI 1640 supplemented with 10% FBS, 1 mM glutamine (Gibco), 1 mM sodium pyruvate (Gibco), 50 μM α-thioglycerol (Sigma-Aldrich), and 20 nM bathocuproine disulfonic acid (BCS) (Sigma-Aldrich). X50-7 and Awia lymphoblastoid cell lines were cultured in RPMI 1640 supplemented with 10% FBS and 1 mM glutamine.
5
Comprehensive Ginsenoside and Amino Acid Analysis
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Ginsenoside Re, Rg1, Rb1, Rb2, Rc, Rd, 20(S)-Rg3, 20(R)-Rg3, 20(S)Rg2, 20(R)-Rg2, 20(S)-Rh1, and 20(R)-Rh1 were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Malonyl ginsenoside Rb1, Rb2, Rc, Rd, zingibroside R1, ginsenoside Ro, Rk1, Rk3, Rg5, Rg6, and F4 were isolated by our laboratory [19] , [20] (link). Arginine, glutamic acid, threonine, asparagine, lysine, phenylalanine, leucine, isoleucine, cysteine, methionine, valine, tyrosine, proline, alanine, histidine, glycine, and serine were purchased from Sigma-Aldrich (Shanghai, China). Methanol and acetonitrile were HPLC grade (Fisher Scientific, Pittsburgh, PA). Phenylisothiocyanate and triethylamine were obtained from Agela of the USA (Agela Technologies Inc., Wilmington, DE).
6
Quantitative Analysis of Amino Acids
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Standards of amino acids (AA) namely phenylalanine (Phe), tryptophan (Trp), isoleucine (Ile), leucine (Leu), asparagine (Asn), methionine (Met), valine (Val), proline (Pro), tyrosine (Tyr), alanine (Ala), threonine (Thr), glycine (Gly), serine (Ser), glutamic acid (Glu), aspartic acid (Asp), arginine (Arg), glutamine (Gln), lysine (Lys), histidine (His), cysteine (Cys) and cystine (Cys2),were purchased from Sigma-Aldrich (Steinheim, Germany). Acetonitrile (ACN) and water (LC-MS grade) was purchased from Carlo Erba (Milan, Italy). Formic acid (LC-MS additive), TGA (thioglycolic acid; ≥98%), and ammonium formate (NH4HCO2) were purchased from Sigma-Aldrich (Steinheim, Germany). Analytical grade hydrochloric acid (HCl) 37% w/w was also supplied from Carlo Erba (Milan, Italy). The chromatographic column HILIC amide BEH, Acquity UPLC 1.7 m, 2 × 150 mm (Waters) were used.
7
Acrylamide Reaction Pathway Chemicals
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Acrylamide (>99.8%), asparagine, glutamine, aspartic acid, alanine, valine, gamma-aminobutyric acid (GABA), NaCl, KCl, MgCl2, CaCl2, thiamine hydrochloride (vitamin B1), nicotinic acid (vitamin B3), pyridoxal hydrochloride (vitamin B6), sucrose, D-glucose, D-fructose, glacial acetic acid (≥99%), and citric acid were obtained from Sigma-Aldrich (St. Louis, MO, USA). Analytical-standard-grade (97%) 5-hydroxymethylfurfural (HMF) was purchased from Thermo Fisher (Kandel GmbH, Kandel, Germany).
8
Comprehensive Analytical Characterization Protocol
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JASCO P-2000 digital polarimeter was used to record the optical rotations while GE Healthcare Ultrospec 9000 spectrophotometer was used to obtain the UV spectra.
Bruker DRX-400 NMR spectrometer with Cryoprobe was used to collect the NMR spectra. 5-mm BBI (1H, G-COSY, multiplicity-edited G-HSQC, and G-HMBC spectra) or BBO (13C spectra) probe heads equipped with z-gradients were used.
Agilent 1260 Infinity Preparative-Scale LC/MS Purification System and Agilent 6130B single quadrupole mass spectrometer for LC and LC/MS Systems were used to perform the preparative HPLC analysis.
Agilent UHPLC 1290 Infinity coupled to Agilent 6540 accurate-mass quadrupole time-of-flight (QTOF) mass spectrometer which was equipped with a splitter and an ESI source were used to acquire the HRESIMS and MS/MS spectra. For over 15 min, under standard gradient condition of 100% water with 0.1% formic acid to 100% acetonitrile with 0.1% formic acid, the analysis was performed with a C18 4.6 × 75 mm, 2.7 μm column at flowrate of 2 mL/min. The operating parameters for QTOF were the same as in [6 (link)].
Nα-(2,4-Dinitro-5-fluorophenyl)-L-alaninamide (L-FDAA) and amino acid standards were purchased from Sigma Aldrich except D-allo-threonine which was purchased from Chem Cruz. Aspartic acid and glutamic acid were converted from asparagine and glutamine (Sigma Aldrich), respectively.
Bruker DRX-400 NMR spectrometer with Cryoprobe was used to collect the NMR spectra. 5-mm BBI (1H, G-COSY, multiplicity-edited G-HSQC, and G-HMBC spectra) or BBO (13C spectra) probe heads equipped with z-gradients were used.
Agilent 1260 Infinity Preparative-Scale LC/MS Purification System and Agilent 6130B single quadrupole mass spectrometer for LC and LC/MS Systems were used to perform the preparative HPLC analysis.
Agilent UHPLC 1290 Infinity coupled to Agilent 6540 accurate-mass quadrupole time-of-flight (QTOF) mass spectrometer which was equipped with a splitter and an ESI source were used to acquire the HRESIMS and MS/MS spectra. For over 15 min, under standard gradient condition of 100% water with 0.1% formic acid to 100% acetonitrile with 0.1% formic acid, the analysis was performed with a C18 4.6 × 75 mm, 2.7 μm column at flowrate of 2 mL/min. The operating parameters for QTOF were the same as in [6 (link)].
Nα-(2,4-Dinitro-5-fluorophenyl)-L-alaninamide (L-FDAA) and amino acid standards were purchased from Sigma Aldrich except D-allo-threonine which was purchased from Chem Cruz. Aspartic acid and glutamic acid were converted from asparagine and glutamine (Sigma Aldrich), respectively.
9
Biochemical Reagents for Cell Culture Experimentation
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Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from HyClone Laboratories, Inc. (Logan, UT, USA). 1% penicillin/streptomycin was obtained from GE Healthcare (Chicago, IL, USA). Dimethyl sulfoxide (DMSO), ethanol, lipopolysaccharides (LPS), nitrite assay kit, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), spermidine, histidine, serine, asparagine, glucose, citrulline, glutamate, glucose-6-phosphate, methionine, itaconate, tryptophan and FMOC-glycine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Lysine, arginine, aspartate, threonine, glutamine, proline, creatine and tyrosine were purchased from Alfa Aesar (Ward Hill, MA, USA). Apigenin, apigetrin, apiin, bergapten and xanthotoxin were purchased from Toronto Research Chemicals (North York, ON, Canada).
10
Comprehensive Metabolite Extraction Protocol
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Methanol, formic acid, ammonium formate, tryptophan, glutamine, asparagine, amino acid standard mixture (AAS18), and isotopically labeled amino acid mix standard (20 amino acids) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Glucosinolates standards including glucocheirolin, progoitrin, glucoraphenin, epiprogoitrin, glucobrassicanapin, glucoalyssin, glucobrassicin, gluconasturtiin, 4-hydroxyglucobrassicin, and glucobarbarin were purchased from Cfm Oskar Tropitzsch GmbH (Marktredwitz, Germany), and sinigrin, glucoiberin, glucotropaeolin, glucoraphanin, gluconapin, glucoerucin, and glucosinalbin were purchased from ChromaDex (Santa Ana, CA, USA). LC-MS grade Acetonitrile was obtained from Merck. In-house purified distilled water was made with a Milli-Q purification system (Bedford, MA, USA).
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