Non-essential amino acids (glycine, 7.5 mg/L; L-alanine, 8.9 mg/L; L-asparagine, 13.2 mg/L; L-aspartic acid, 13.3 mg/L; L-glutamic acid, 14.7 mg/L; L-proline, 11.5 mg/L; and L-serine, 10.5 mg/L) and sodium pyruvate (1 mM) were purchased from Life Technologies. The apoptosis inhibitor, M5054 [100 μg/mL; 2,2′-methylenebis(1,3-cyclohexanedione)], was purchased from Merck (Billerica, MA, USA). 2-(N-(5-chloro-2-methylphenyl)methylsulfonamido)-N-(2,6-difluorophenyl)acetamide), (10 nM; hepatocyte functional proliferation enhancer, FPH1) was purchased from XcessBio (San Diego, CA, USA) [23 (link)]. Galactose (900 mg/mL), ornithine (1 mM), oncostatin M (20 ng/mL), nicotinamide (1.2 mg/mL), proline (30 ng/mL), and L-glutamine (0.3 mg/mL) were purchased from Wako Pure Chemicals (Osaka, Japan).
Proline
Manufactured by Fujifilm
Sourced in Japan
The Proline is a line of lab equipment designed and manufactured by Fujifilm. It serves as a versatile platform for various laboratory applications. The Proline provides reliable and consistent performance to support the needs of research and scientific professionals.
Automatically generated - may contain errors
Lab products found in correlation
Tyrosine, by Fujifilm (3 mentions)
Histidine, by Fujifilm (3 mentions)
Phenylalanine, by Fujifilm (3 mentions)
Galactose, by Fujifilm (2 mentions)
Ornithine, by Fujifilm (2 mentions)
Glycine, by Fujifilm (2 mentions)
Glutamine, by Fujifilm (2 mentions)
Cysteine, by Fujifilm (2 mentions)
Alanine, by Fujifilm (2 mentions)
Arginine, by Fujifilm (2 mentions)
Asparagine, by Fujifilm (2 mentions)
Aspartic acid, by Fujifilm (2 mentions)
Glutamic acid, by Fujifilm (2 mentions)
Serine, by Fujifilm (2 mentions)
Isoleucine, by Fujifilm (2 mentions)
Leucine, by Fujifilm (2 mentions)
Lysine, by Fujifilm (2 mentions)
Methionine, by Fujifilm (2 mentions)
Threonine, by Fujifilm (2 mentions)
Tryptophan, by Fujifilm (2 mentions)
8 protocols using proline
1
Hepatocyte Proliferation Enhancing Protocol
2
Amino Acid Quantification Protocol
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All of the amino acids (alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, serine, tyrosine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, ornithine, and citrulline), pyroglutamic acid, urocanic acid, urea, and lactic acid were purchased from Wako (Osaka, Japan). All of the SIL-IS were purchased from JUNSEI (Tokyo, Japan). Deionized water was produced in-house using a Milli-Q gradient water purification system (Merck Millipore, Darmstadt, Germany). The SIL-IS mixture solution containing all of the SIL compounds was prepared in deionized water with each having a final concentration of 0.5 μmol/mL. The NMF mixture solutions containing all of the amino acids and pyroglutamic acid, urocanic acid, urea, lactic acid were prepared in deionized water with a final concentration ranging between 0.025–12.5 μmol/mL for each compound.
3
Chondrogenic Differentiation of MSCs
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MSCs (1 × 106) were placed in polypropylene tubes and centrifuged at a speed of 300 × g for 6 min. The pellets were cultured in 1 ml complete chondrogenic medium consisting of Dulbecco’s modified Eagle’s medium (Life Technologies, Gaithersburg, MD, USA) supplemented with 10−7 M dexamethasone (Sigma), 1% insulin-transferrin-sodium selenium (Sigma), 50 μM ascorbate-2-phosphate (WAKO), 50 μg/ml proline (WAKO), 20 ng/ml transforming growth factor-β3 (Life Technologies), and 1% antibiotics (WAKO)39 (link). The polypropylene tubes were maintained at 37 °C in a 5% CO2 incubator, and the medium was changed every 3 days. The pellets were used for immunostaining or qPCR at each time. Data were normalized to the average mRNA level at day 0 (set at 1) and are presented as means ± SEM.
4
Rice Cultivar Salt Stress Response
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Seeds of two rice (Oryza sativa L.) cultivars, cv. BRRI dhan49 (salt sensitive) and cv. BRRI dhan54 (salt tolerant) were collected from Bangladesh Rice Research Institute (BRRI) and surface-sterilized with 70% ethanol for 10 min followed by washing several times with sterilized distilled water. Seeds were then soaked for 24 h in the dark. Seeds were sown on plastic nets upon plastic beakers containing distilled water and kept in the dark at 28 ± 2°C for germination. After 48 h, uniformly germinated seeds were transferred to growth chamber with controlled conditions (light intensity, 100 μmol m−2 s−1; temperature, 25 ± 2°C; relative humidity, 65–70%) and during the growing period hyponex solution (Hyponex, Japan) was used as nutrient. Two levels of salt stresses (150 and 300 mM NaCl) were imposed on fourteen-day-old rice seedlings with, without 5 mM proline [l (-) proline, Wako, Japan] and betaine (betaine, Wako, Japan), and where these protectants were sprayed twice a day mixing with the wetting agent 0.02% Tween 20 (Tween 20, Wako, Japan). Control plants were grown with Hyponex solution only. Data were taken after 48 h of NaCl treatment. The experiment was repeated three times (n = 3) under the same conditions.
5
Defined Xeno-Free Hepatocyte Culture Protocol
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HSM was prepared from amino acid powders following the formulation of Leibovits-15 medium (Life Technologies) [18 (link)]. This HSM lacked arginine, tyrosine, glucose, and sodium pyruvate, but was supplemented with galactose (900 mg/L), ornithine (1 mM), glycerol (5 mM), and proline (260 mM) (all from Wako Pure Chemicals). proline (30 mg/L) was added for DNA synthesis to occur [21 (link)]. Aspartic acid was not included since it is one of the products of ornithine and an arginine substrate. KSR (Life Technologies) was added at a final concentration of 10%, and used instead of fetal calf serum to establish defined xeno-free conditions.
HDI was prepared by mixing oncostatin M (20 ng/mL), FPH1, M50054 (100 μg/mL), non-essential amino acids (glycine, 7.5 mg/L; L-alanine, 8.9 mg/L; L-asparagine, 13.2 mg/L; L-aspartic acid, 13.3 mg/L; L-glutamic acid, 14.7 mg/L; L-proline, 11.5 mg/L; and L-serine, 10.5 mg/L), sodium pyruvate (1 mM), nicotinamide (1.2 mg/mL), proline (30 ng/mL), and glutamine (0.3 mg/mL). proline and nicotinamide are necessary for primary hepatocytes to proliferate [21 (link), 22 (link)].
HDI was prepared by mixing oncostatin M (20 ng/mL), FPH1, M50054 (100 μg/mL), non-essential amino acids (glycine, 7.5 mg/L; L-alanine, 8.9 mg/L; L-asparagine, 13.2 mg/L; L-aspartic acid, 13.3 mg/L; L-glutamic acid, 14.7 mg/L; L-proline, 11.5 mg/L; and L-serine, 10.5 mg/L), sodium pyruvate (1 mM), nicotinamide (1.2 mg/mL), proline (30 ng/mL), and glutamine (0.3 mg/mL). proline and nicotinamide are necessary for primary hepatocytes to proliferate [21 (link), 22 (link)].
6
Spectroscopic Analysis of Amino Acid Interactions
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Asparagine, glycine, alanine, arginine, aspartic acid, cysteine, glutamine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, valine, Trp, 10% hydrochloric acid (HCl), sodium hydroxide (NaOH), ascorbic acid (AA), sodium chloride (NaCl), D (+)-glucose, lysozyme, and glyoxylic acid monohydrate were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). MSG monohydrate and NaOCl·5H2O were purchased from Nacalai Tesque (Kyoto, Japan). Pooled Human Cerebrospinal Fluid (CSF) was purchased from Lee Biosolutions, Inc. (USA, Missouri).
A block incubator, WSC-2620 PowerBLOCK (ATTO Corporation, Tokyo, Japan), was used to set and maintain the temperature (10, 20, 30, 40, 50, 60, and 70°C) of sample solutions.
A double-beam spectrophotometer U-2900 (Hitachi High-Technology Co., Ltd., Tokyo, Japan) with microcells and a 10-mm path length was used to measure the absorption spectra of sample solutions.
A block incubator, WSC-2620 PowerBLOCK (ATTO Corporation, Tokyo, Japan), was used to set and maintain the temperature (10, 20, 30, 40, 50, 60, and 70°C) of sample solutions.
A double-beam spectrophotometer U-2900 (Hitachi High-Technology Co., Ltd., Tokyo, Japan) with microcells and a 10-mm path length was used to measure the absorption spectra of sample solutions.
7
Genetic Manipulation of E. coli Strains
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The strains used in this study are listed in Table S3 . Bacterial cells were grown at 37℃ with reciprocal shaking at 180 rpm in an LB Miller medium (BD Biosciences) or an M9 minimal medium supplemented with 0.2% glucose, 10 µg/ml thiamine (Fujifilm Wako Pure Chemical), and 200 µg/ml proline (Fujifilm Wako Pure Chemical). Where required, media were supplemented with antibiotics at the following concentrations: 50 µg/ml ampicillin (Ap), 50 µg/ml kanamycin (Km), and 12.5 µg/ml chloramphenicol (Cm).
Deletion strains were constructed using the lambda Red system as previously described (Datsenko & Wanner, 2000 (link)). E. coli gcvB, sroC, dppABCDF, and ydeE were deleted by chromosomal insertion of the pKD4‐ or pKD13‐derived PCR fragments amplified with primer pairs JVO‐0131/JVO‐0132, JVO‐7614/JVO‐7615, dppA‐P1‐R/dppA‐P4‐F, and ydeE‐P1‐R/ydeE‐P4‐F, respectively (TableS4 ). The 3xFLAG epitope tag at the C‐terminus of gdhA was amplified with the primer pair MMO‐0206/MMO‐0207 using pSUB11 (Uzzau et al., 2001 (link)) as a template and was introduced into the chromosome using the lambda Red system (Datsenko & Wanner, 2000 (link)). The deletion or insertion in the chromosome was confirmed by PCR, and the mutant loci were moved into the appropriate strains by P1 phage‐mediated transduction.
Deletion strains were constructed using the lambda Red system as previously described (Datsenko & Wanner, 2000 (link)). E. coli gcvB, sroC, dppABCDF, and ydeE were deleted by chromosomal insertion of the pKD4‐ or pKD13‐derived PCR fragments amplified with primer pairs JVO‐0131/JVO‐0132, JVO‐7614/JVO‐7615, dppA‐P1‐R/dppA‐P4‐F, and ydeE‐P1‐R/ydeE‐P4‐F, respectively (Table
8
Lanthanide-Based Affinity Ligand Synthesis
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All reagents employed were of analytical grade. Neu5Ac and disialic acid (diNeu5Ac) were purchased from Nagara Science (Gifu, Japan). The affinity ligands, S-2-(4-aminobenzyl)-1,4,7triazacyclononane-1,4,7-triacetic acid (ABNOTA) and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) were purchased from Macrocyclics (Dallas, TX). Chloride salts of lanthanides (lanthanum, europium, terbium, dysprosium, erbium, ytterbium and lutetium) were obtained from Sigma-Aldrich (St. Louis, MO). The pH buffer reagent, N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), was employed (Dojindo, Kumamoto, Japan). Dimethyl sulfoxide (DMSO), glycine, proline, phenylalanine, tryptophan, histidine and tyrosine were purchased from Wako Pure Chemical Industry (Osaka, Japan). D-fructose, D-glucose and D-ribose were purchased from Kanto Kagaku (Tokyo, Japan). All stock solutions were prepared by dissolving appropriate amounts of the solid reagents into ultrapure water (>18.2 MΩ•cm), produced by Direct-Q 3UV (Nihon Millipore, Tokyo, Japan).
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