L-Thr, L-Ser, dithiothreitol (DTT), ribonucleoside triphosphate (NTP), guanosine monophosphate (GMP), tetrasodium pyrophosphate, pyrophosphatase (PPiase), Tris-base, MgCl2, NaCl, activated charcoal, horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Sigma (St Louis, MO, USA). [14C]Thr, [14C]Ser and [α-32P]ATP were obtained from Perkin Elmer Inc. (Waltham, MA, USA). KOD-plus mutagenesis kits, ReverTra Ace quantitative polymerase chain reaction (qPCR) RT Kit and SYBR Green Realtime PCR Master Mix were obtained from TOYOBO (Osaka, Japan). Dynabeads protein G, Lipofectamine 2000 transfection reagent, and the protein synthesis kit (1-Step Human Coupled IVT Kit) were obtained from Thermo Scientific (Waltham, MA, USA). Qproteome nuclear protein kits and Ni2+-NTA Superflow resin were purchased from Qiagen Inc. (Hilden, Germany). Polyethyleneimine cellulose plates were purchased from Merck (Darmstadt, Germany).
L thr
Manufactured by Merck Group
Sourced in United States
L-Thr is a type of laboratory equipment used for the detection and analysis of L-threonine, an essential amino acid. The core function of this product is to provide accurate and reliable measurements of L-threonine concentrations in various samples.
Automatically generated - may contain errors
Lab products found in correlation
L ser, by Merck Group (5 mentions)
Kod plus mutagenesis kit, by Toyobo (3 mentions)
Sodium chloride (nacl), by Merck Group (3 mentions)
Mgcl2, by Merck Group (3 mentions)
Tris base, by Merck Group (3 mentions)
Ni2 nta superflow resin, by Qiagen (3 mentions)
L ala, by Merck Group (3 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Merck Group (2 mentions)
Polyethyleneimine cellulose plate, by Merck Group (2 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
α 32p atp, by PerkinElmer (2 mentions)
Tetrasodium pyrophosphate, by Merck Group (2 mentions)
T4 dna ligase, by Thermo Fisher Scientific (2 mentions)
Pyrophosphatase ppiase, by Roche (2 mentions)
L tyr, by Merck Group (2 mentions)
L leu, by Merck Group (2 mentions)
L val, by Merck Group (2 mentions)
L pro, by Merck Group (2 mentions)
1 step human coupled ivt kit, by Thermo Fisher Scientific (1 mentions)
7 protocols using l thr
1
In Vitro Protein Synthesis Assay
2
Enzymatic Synthesis and Purification of GSNO
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L-Thr, L-Ser, NTP (CTP, ATP, UTP and GTP), GMP, Tris-base, MgCl2, NaCl, dithiothreitol (DTT), and tetrasodium pyrophosphate were purchased from Sigma (St. Louis, MO, USA). Pyrophosphatase (PPiase) was obtained from Roche Life Science (Shanghai, China). [32P]tetrasodium pyrophosphate (NEX019001MC) and [α-32P]ATP (BLU503H250UC) were purchased from PerkinElmer (Shelton, CT, USA). KOD-Plus mutagenesis kits were obtained from TOYOBO (Osaka, Japan). All restriction endonucleases, T4 DNA ligase, and T4 polynucleotide kinase were obtained from Thermo Scientific (Waltham, MA, USA). Ni2+-NTA Superflow resin was purchased from Qiagen Inc. (Hilden, Germany). DNA sequencing and primer synthesis were performed by Biosune (Shanghai, China). GSNO was synthesized from glutathione with acidified nitrite as described previously (38 (link)). Briefly, GSH reacted with the equimolar sodium nitrite at 4°C in HCl (625 mM) for 45 min in the dark. Then the 2.5 volumes of acetone were added and the mixture was stirred constantly for 20 min. GSNO was washed by acetone and dried under vacuum. Prepared GSNO was quantified by the absorbance of 334 nm.
3
Reagents for Molecular Biology Experiments
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l -Thr, l -Ser, dithiothreitol, nucleoside triphosphates (NTPs), guanosine monophosphate (GMP), tetrasodium pyrophosphate, Tris-base, MgCl2, NaCl and inorganic pyrophosphate were purchased from Sigma (St Louis, MO, USA). [14C]Thr was obtained from Biotrend Chemicals (Destin, FL, USA). [α-32P]ATP and [γ-32P]ATP were obtained from Perkin Elmer Inc. (Waltham, MA, USA). T4 DNA ligase, T4 PNK (polynucleotide kinase), RNase T1, RNase S1, and restriction endonucleases were obtained from Thermo Scientific (Pittsburgh, PA, USA). Phusion high-fidelity DNA polymerase was purchased from New England Biolabs (Ipswich, MA, USA). Ni2+-NTA (nitrilotriacetic acid) Superflow was purchased from Qiagen Inc. (Germany). Pyrophosphatase (PPiase) was obtained from Roche Applied Science (China). The dNTP mixture was purchased from Takara (Japan). Oligonucleotide primers were synthesized by Biosune (China). Escherichia coli Rosetta (DE3) cells were purchased from Stratagene (Santa Clara, CA, USA).
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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
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).
6
Purification and Characterization of Threonine Kinase
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l -Thr, NTP, GMP, tetrasodium pyrophosphate, pyrophosphatase (PPiase), Tris-base, MgCl2, MnCl2, NaCl, DTT, NaHCO3, activated charcoal, anti-FLAG (F7425), anti-GAPDH (G8795) antibodies, horseradish peroxidase (HRP)-conjugated secondary antibodies, standard proteins (including bovine serum albumin, ovalbumin, carbonic anhydrase, ribonuclease A and aprotinin) and biotinamidohexanoic acid hydrazide (B3770-25MG) were purchased from Sigma (St. Louis, MO, USA). Anti-Myc (HOA012MC), anti-HA (HOA012HA) and anti-His6 (HOA012HS) were purchased from Shanghai HuiOu Biotechnology Co. Ltd (Shanghai, China). [α-32P]ATP, [14C]Thr was obtained from Perkin Elmer Inc. (Waltham, MA, USA). KOD-plus mutagenesis kits were obtained from TOYOBO (Osaka, Japan). Yeast was transformed using a Yeastmaker Yeast Transformation System 2 kit (Takara Bio, Japan). Lipofectamine 2000 transfection reagent, SuperSignal West 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). Primer synthesis and DNA sequencing were performed by Biosune (Shanghai, China).
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7
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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