1,8-Diaminonaphthalene, m-vanillin, methanol, l -Ala, l -Arg, l -asparagine (l -Asp), l -cysteine (l -Cys), glucose (GOx), hydrogen peroxide (H2O2), ascorbic acid (AA), dopamine (DA), anhydrous potassium carbonate (K2CO3), hexane, tetrahydrofuran (THF) and N,N′-dicyclohexylcarbodiimide (DCC), dimethyl sulfoxide (DMSO) were supplied by Sigma Aldrich. N,N′-Dimethylformamide (DMF) was obtained from M-Tedia (USA) and used without further purification.
L ala
Manufactured by Merck Group
Sourced in United States
L-Ala is a laboratory equipment product. It is a type of amino acid commonly used in various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
L leu, by Merck Group (4 mentions)
L ser, by Merck Group (4 mentions)
L thr, by Merck Group (3 mentions)
L arg, by Merck Group (2 mentions)
L gly, by Merck Group (2 mentions)
L glu, by Merck Group (2 mentions)
L asp, by Merck Group (2 mentions)
L tyr, by Merck Group (2 mentions)
L val, by Merck Group (2 mentions)
L met, by Merck Group (2 mentions)
L pro, by Merck Group (2 mentions)
L ile, by Merck Group (2 mentions)
1 8 diaminonaphthalene, by Merck Group (1 mentions)
Tetrahydrofuran thf, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
L 3 4 3h n glutamine, by PerkinElmer (1 mentions)
Wallac 1409 liquid scintillation counter, by PerkinElmer (1 mentions)
Standard proteins, by Merck Group (1 mentions)
Tetrasodium pyrophosphate, by Merck Group (1 mentions)
Polyethyleneimine cellulose plate, by Merck Group (1 mentions)
9 protocols using l ala
1
Colorimetric Dopamine Sensing Assay
2
Astrocyte [3H]glutamine Uptake Assay
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Astrocytes were washed twice with Krebs buffer (29.5 mM NaCl, 1.13 mM KCl, 0.3 mM KH2PO4, 0.3 mM MgSO4, 11 mM glucose, 25 mM NaHCO3, 2.5 mM CaCl2) and pre-incubated in this buffer for 15 min at 37 °C. Next, the cells were incubated in the mixture of Krebs buffer supplemented with 0.1 μCi/mL L-[3,4-3H(N)-]glutamine (PerkinElmer, Waltham, MA, USA; specific radioactivity 37 MBq/mL) and 0.1 mM unlabeled glutamine. To block systems other than system N in the experiments analyzing system N-mediated [3H]glutamine uptake, the mixture also contained 10 mM L-Ala and 10 mM L-Leu (Sigma-Aldrich, St. Louis, MO, USA) [9 (link)]. The incubation was terminated after 4 min by a triple wash of the cells using cold Krebs buffer. The astrocytes were lysed by incubation with 0.5 mL of 1 N NaOH and the radioactivity of the cells was measured in a Wallac 1409 Liquid Scintillation Counter (Perkin-Elmer, Turku, Finland).
3
Affinity Purification of Recombinant Proteins
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L-Ala, L-Ser, L-Gly, NTP, GMP, tetrasodium pyrophosphate, pyrophosphatase (PPiase), Tris-base, MgCl2, NaCl, activated charcoal, anti-FLAG (F7425) antibodies (the FLAG tag epitope has the sequence motif N-DYKDDDDK-C), horseradish peroxidase-conjugated secondary antibodies, standard proteins (including apoferritin from horse spleen, yeast alcohol dehydrogenase and bovine serum albumin) were purchased from Sigma (St. Louis, MO, USA). [14C]Ala and [α-32P]adenosine triphosphate (ATP) were obtained from Perkin Elmer Inc. (Waltham, MA, USA). KOD-plus mutagenesis kits were obtained from TOYOBO (Osaka, Japan). Lipofectamine 2000 and 3000 transfection reagents and Dynabeads protein G were obtained from Thermo Scientific (Waltham, MA, USA). Ni2+-NTA Superflow resin was purchased from Qiagen Inc. (Hilden, Germany). Polyethyleneimine cellulose plates were purchased from Merck (Darmstadt, Germany).
4
Synthesis of Ruthenium Nitrosyl Complexes
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The starting compounds Na2[RuCl5NO]·6H2O and (nBu4N)2[RuCl5NO] were synthesized as previously
reported in the literature.24 RuCl3·H2O was purchased from Johnson Matthey, sodium
nitrite (97%), tetrabutylammonium chloride (97%),l -Thr, l -Ala, and Gly (99%) were from Sigma-Aldrich. l -Ser
was from Serva,l -Pro (99%), and d -Pro (99%) were
from Alfa Aesar, andl -Tyr (99%), formic acid, and sodium
ascorbate were from Fluka. All chemicals were used without further
purification. Methanol (HPLC grade, Fisher) and ultrapure water (18.2
MΩ, Advantage A10, 185 Ultrapure Water System, Millipore, France)
were used for the ESI-MS study.
reported in the literature.24 RuCl3·H2O was purchased from Johnson Matthey, sodium
nitrite (97%), tetrabutylammonium chloride (97%),
was from Serva,
from Alfa Aesar, and
ascorbate were from Fluka. All chemicals were used without further
purification. Methanol (HPLC grade, Fisher) and ultrapure water (18.2
MΩ, Advantage A10, 185 Ultrapure Water System, Millipore, France)
were used for the ESI-MS study.
5
Metabolic Tracing of Amino Acids
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The following were purchased from Sigma-Aldrich: L-Ala (A7469), L-Asn (A4159), L-Asp (A7219), L-Glu (G8415), L-Ser (S4311), L-Gln (G8540), L-Arg (A8094), L-Cys (C7352), L-Pro (P5607), NH4Cl (A9434), dmKG (349631), L-cycloserine (C1159) at 250 μM, DEN (N-0756), and GDH (G2626). Also used were rapamycin (LC Laboratories, R-5000; 2 mg/kg for in vivo, 10 μM for in vitro); β-CLA (Santa Cruz Biotechnology, sc-291972) at 250 μM for cell culture, and 20 mg/kg for in vivo experiments; and DL-Cyc (MCE, HY-W008440) at 250 μM. Direct Red 80 stable isotope tracers were purchased from Cambridge Isotope Laboratories: 15N-NH4Cl (NLM-467-PK), U-13C-D-glucose (CLM-1396-1), 15N-NH4OAc (NLM-177-PK), α-15N-L-Gln (NLM-1016-PK), amide-15N-L-Gln (NLM-557-PK), and U-13C-aKG (CLM-2411-PK).
6
Biogenic Amine and Amino Acid Analysis
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BA standards (tyramine hydrochloride, 2-phenylethylamine hydrochloride, putrescine dihydrochloride, cadaverine dihydrochloride, histamine dihydrochloride, tryptamine hydrochloride, spermidine trihydrochloride) and amino acid standards (L-Asp, L-Glu, L-Ser, L-Gly, L-His, L-Thr, L-Arg, γ-aminobutyric acid [GABA], L-Ala, L-Pro, L-Tyr, L-Val, L-Met, L-Iso, L-Leu, L-Phe, L-Trp, and L-Lys) were all purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Dansyl chloride, perchloric acid, sodium hydrogen carbonate, potassium dichromate, and silver nitrate were purchased from Daejung Chemical Co. (Siheung, Korea). Distilled water, acetone, and acetonitrile (HPLC grade) were purchased from Tedia Co. (Fairfield, OH, USA).
7
Gel Preparation and Specimen Staining
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AgarArt powder (CTS S.r.l., Milan, Italy) was used for gel preparation. Disodium EDTA (Merck-Millipore, Darmstadt, Germany), TAC (Bresciani S.r.l., Milan, Italy) and L-ALA (Sigma Aldrich, CAS n. 56-41-7, Darmstadt, Germany) were used as chelating agents. CuSO4·5H2O (Sigma Aldrich, Darmstadt, Germany) was used as a copper source. For laboratory specimen staining, Na2CO3 (Merck-Millipore, Darmstadt, Germany) was also used. For pH solution modification, (NH4)2CO3, H2SO4 97% and NH3 33% (Sigma Aldrich, Darmstadt, Germany) were used. HNO3 (70%, Carlo Erba, Cornaredo, Italy) was used for gel dissolution. MQ water® and ultra-pure water were used when necessary.
8
Amino Acid Pretreatment on Plant Immunity
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L -Ala (A7469, Sigma-Aldrich, United States), L -Leu (L8912, Sigma-Aldrich, United States), L -Ile (I2752, Sigma-Aldrich, United States), L -Met (M5308, Sigma-Aldrich, United States), L -Phe (V900489, VETEC, United States), L -Pro (V900338, VETEC, United States), L -Trp (V900470, VETEC, United States), L -Val (V900465, VETEC, United States), L -Asn (900458, VETEC, United States), L -Cys (V900400, VETEC, United States), Gly (V900144, VETEC, United States), L -Gln (V900419, VETEC, United States), L -Ser (V900406, VETEC, United States), L -Thr (V900466, VETEC, United States), L -Tyr (V900426, VETEC, United States), L -Asp (V900407, VETEC, United States), L -Glu (V900408, VETEC, United States), L -Arg (V900343, VETEC, United States), L -His (V900459, VETEC, United States), and L -Lys (V9 00409, VETEC, United States) were dissolved in 0.05% (v/v) Tween 20 solution to prepare amino acid solutions at a certain concentration. Plants grown in a room under a 10-h light/14-h dark photoperiod (21–23°C) were sprayed with amino acid solution or 0.05% Tween 20 solution (Mock), and then continued to grow for another 2 days. Subsequently, the pretreated plants were inoculated with B. cinerea spore suspension.
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9
Amino Acid Analysis of Sclerin and Cyclosenegalin A
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Sclerin (1 ) (200 μg) and cyclosenegalin A (2 ) (200 μg) were hydrolyzed in 200 μL 6 M HCl at 50 °C for 18 h. After, the residual HCl was removed in vacuo, and then 100 μL of an acetone solution containing 0.1 M of NaHCO3 and 25 μg of 1-fluoro-2,4-dinitrophenyl-5-l -alaninamide (l -FDAA) was added to the residue. The solution mixture was heated at 75 °C for 4 h. Next, the reaction mixture was cooled, neutralized with 2 M HCl (50 µL) and dissolved in MeOH (200 μL). About 10 µL of each solution of FDLA derivatives was analyzed by HPLC. On the other hand, authentic standards of l -Dab, l -Pro, l -Ile, l -Leu, l -Ala, l -Val, l -Thr, l -Tyr, l -Ser (Sigma-Aldrich, St Louis, MO, USA) were treated with l -FDAA, as described above. The l -FDAA derivative of l -amino acid standard were analyzed by HPLC–UV, and the retention times of l -Dab (2.6), l -Pro (4.2), l -Ile (20.1), l -Leu (5.5), l -Ala (4.7), l -Val (9.8), l -Thr (3.0), l -Tyr (6.4), l -Ser (4.6) were compared with the Marfey’s derivative of 1 and 2 . HPLC conditions: a 5 μM column X-Terra MS C-18 (150 × 3.0 mm) maintained at 25 °C was eluted at 1 mL/min with 40% MeOH/H2O containing 0.01% HCOOH for 25 min.
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