L ascorbate

Manufactured by Merck Group

Sourced in United States, Germany

L-ascorbate is a laboratory product that functions as a reducing agent in various biochemical and analytical applications. It is the salt or ester form of ascorbic acid, also known as vitamin C. L-ascorbate is commonly used in cell culture media, redox reactions, and as a stabilizer in various solutions.

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Lab products found in correlation

β glycerophosphate, by Merck Group (8 mentions) Dexamethasone (dex), by Merck Group (3 mentions) Alizarin red, by Merck Group (3 mentions) Streptomycin, by Merck Group (3 mentions) Penicillin, by Merck Group (3 mentions) Aqua bidest, by Merck Group (2 mentions) Alpha mem, by Thermo Fisher Scientific (2 mentions) Osteo assay surface plate, by Corning (2 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions) Collagenase, by Merck Group (2 mentions) Dispase 2, by Merck Group (2 mentions) Menadione, by Merck Group (2 mentions) Alizarin red s, by Merck Group (2 mentions) Sodium pyruvate, by Merck Group (1 mentions) 3 isobutyl 1 methylxanthine, by Merck Group (1 mentions) Insulin, by Merck Group (1 mentions) Indomethacin, by Merck Group (1 mentions) Transforming growth factor β1, by Thermo Fisher Scientific (1 mentions) Alizarin red s, by Cyagen (1 mentions) Alcian blue, by Cyagen (1 mentions)

Spelling variants of this product

Ascorbate (130 citations) Sodium L-ascorbate (90 citations) L-ascorbate-2-phosphate (36 citations) Calcium l-ascorbate dihydrate (3 citations) L‐ascorbate acid (2 citations) L-ascorbate-2-phospate (2 citations) (+)-Sodium l-ascorbate BioXtra (2 citations) Vitamin C (Sodium L-ascorbate) (2 citations)

24 protocols using l ascorbate

Multilineage Differentiation of Stem Cells

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hAECs (passages 2) and hAMSCs (passage 3) were tested for their ability to differentiate into osteocytes, chondrocytes and adipocytes.
To induce differentiation into osteocytes, the cells were cultured in osteocyte differentiation medium: 1 μM dexamethasone, 50 μg/ml L-ascorbate, and 10 mM β-glycerophosphate (Sigma-Aldrich) in DMEM supplemented with 10% FBS. After 14 days of differentiation, the cells were fixed and stained with Alizarin Red S (Cyagen, Guangzhou, China).
To induce differentiation into adipocytes, the cells were cultured with adipocyte differentiation medium: 0.5 mM 3-isobutyl-1-methyl xanthine, 1 μM dexamethasone, 200 μM indomethacin, and 10 μg/ml insulin (Sigma-Aldrich) in DMEM supplemented with 10% FBS. After 14 days of differentiation, the cells were stained with Oil Red O (Cyagen).
To induce differentiation into chondrocytes, the cells were cultured with chondrocyte differentiation medium: 0.1 μM dexamethasone, 50 μg/ml L-ascorbate, 100 μg/ml sodium pyruvate (Sigma-Aldrich), and 10 ng/ml transforming growth factor (TGF)-β1 (PeproTech) in DMEM supplemented with 10% FBS. After 14 days of differentiation, the cells were stained with Alcian blue (Cyagen).

Wu Q., Fang T., Lang H., Chen M., Shi P., Pang X, & Qi G. (2017). Comparison of the proliferation, migration and angiogenic properties of human amniotic epithelial and mesenchymal stem cells and their effects on endothelial cells. International Journal of Molecular Medicine, 39(4), 918-926.

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Quantifying Iron Utilization Rates

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The iron utilization rates of M5-90, the irr mutant and the rirA mutant in normal TSB, iron-deficient TSB, or iron-sufficient TSB were measured via 1, 10-phenanthroline chelation with ferrous iron, yielding an orange-red color complex with maximum absorption at 512 nm [38 (link)]. Briefly, during the growth kinetics assay of these three strains, 2 mL cell suspension of M5-90, irr mutant and rirA mutant from the different iron concentrations of TSB was collected at each time point and centrifuged at 12,000 × g for 5 min. The supernatant was transferred into a new tube, and 600 μL of 1, 10-phenanthroline (5 nM, Sigma-Aldrich) and 400 μL of L-ascorbate (0.5 mM, Sigma-Aldrich) were added and incubated for 2 h at 25°C. Finally, the absorbance value was measured at 512 nm wavelength with a Nanodrop 2000 spectrophotometer (Thermo, USA).

Zhang H., Sun T., Cao X., Wang Y., Ma Z., Wang Y., Yang N., Xu M., Deng X., Li H., Wang B., Yi J., Wang Z., Zhang Q, & Chen C. (2023). Scanning iron response regulator binding sites using Dap-seq in the Brucella genome. PLOS Neglected Tropical Diseases, 17(7), e0011481.

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Antioxidant Activity Evaluation Protocol

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Gallic acid (GAE), quercetin (QE), 1,1-diphenyl-2-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), L-ascorbate, Folin-Ciocalteu’s reagent, sodium carbonate, sodium acetate, and aluminum chloride were purchased from Sigma-Aldrich (St Louis, MO, USA). Quercetin was purchased from TCI (Tokyo, Japan). Caffeine was purchased from Sigma-Aldrich (Buchs, Switzerland). Acetonitrile LC-MS grade was purchased from Labscan, (Bangkok, Thailand). Ultrapure water was prepared by using a millipore MilliQ Integral water purification system (Millipore, Bedford, MA, USA). Formic acid (analytical grade) was obtained from Merck (Darmstadt, Germany). Analytical grade of 95% ethanol was purchased from CHEMIPAN (Bangkok, Thailand).
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Oil Red O reagent and human recombinant insulin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dexamethasone (DEX) and isobutylmethylxanthine (IBMX) were purchased from Merck (Kenilworth, NJ, USA). The other chemicals were analytical grade. Bovine calf serum (BCS), fetal bovine serum (FBS), high-glucose Dulbecco’s modified Eagle’s medium (DMEM), phosphate-buffered saline (PBS), trypsin-EDTA and antibiotics (P/S) were purchased from GIBCO (Grand Island, NY, USA).

Ngamdokmai N., Ingkaninan K., Scholfield C.N., Insumrong K., Neungchamnong N., Minale G, & Warinhomhoun S. (2022). A Thai Traditional Triple-Fruit Formulation “Phikud Tri-Phon” May Provide Fat Loss and Nutritional Benefits. Foods, 11(19), 3067.

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Osteoclast and Osteoblast Generation Protocol

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Osteoclast generation was done as previously described (Saferding et al., 2017).
For bone resorption assays, osteoclasts were seeded 600,000 cells/well on osteo‐assay surface plates (Corning) and cultured as previously described (Saferding et al., 2017). After 4 days, osteoclasts were removed using 5% sodium hypochlorite for 5 min and washed two times with water, water was removed, and wells were air‐dried.
Osteoblasts were isolated from calvariae of neonatal WT and miR‐146a^−/−mice. Calvariae were digested for 10 min in alpha MEM (Gibco) containing 0.1% Collagenase (Sigma) and 0.2% Dispase II (Sigma). Osteogenic precursors were expanded in alpha MEM containing 10% FCS (Gibco) and 1% penicillin /streptomycin. Differentiation of osteoblasts was achieved by adding 0.2 mM l‐ascorbate and 10 mM β‐glycerophosphate (both Sigma). After 15, 21, and 26 days, cells were fixed with 4% formalin, following staining with either Aqua Bidest containing 8% Alizarin Red (Sigma) or staining for alkaline phosphatase using 5‐bromo‐4‐chloro‐3‐indolylphosphate/nitro blue.

Saferding V., Hofmann M., Brunner J.S., Niederreiter B., Timmen M., Magilnick N., Hayer S., Heller G., Steiner G., Stange R., Boldin M., Schabbauer G., Weigl M., Hackl M., Grillari J., Smolen J.S, & Blüml S. (2020). microRNA‐146a controls age‐related bone loss. Aging Cell, 19(11), e13244.

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Oxidative Stress and Excitotoxicity Assays

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potassium superoxide (KO₂) (Sigma), dimethyl sulfoxide (DMSO) (Sigma), 18-crown-6 (Sigma), bilirubin IXα (Frontier Scientific), biliverdin IXα (Sigma), bilirubin ditaurate (Lee Biosciences), hemin (Frontier Scientific), pyrogallol (Sigma), paraquat (Santa Cruz Biotech), menadione (Sigma), hydrogen peroxide (H₂O₂) (Sigma), 4-hydroxynonenal (4-HNE) (Cayman Chemical), methylthiazoletetrazolium (MTT) (Sigma), rotenone (Sigma), dihydroethidium (DHE) (Thermo Fisher Scientific), MitoSOX Red (Thermo Fisher Scientific), Hoechst 33258 (Abcam), p-nitro blue tetrazolium (Sigma), 5,5-dimethyl-1-pyrroline N-oxide (DMPO) (Cayman Chemical), L-glutathione (Sigma), L-cysteine (Sigma), L-ascorbate (Sigma), (S)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (S-AMPA) (Tocris Bioscience), kainic acid (Tocris Bioscience), N-Methyl-D-aspartic acid (NMDA) (Sigma), (+)-MK-801 (Sigma), and glycine (Sigma)

Vasavda C., Kothari R., Malla A.P., Tokhunts R., Lin A., Ji M., Ricco C., Xu R., Saavedra H.G., Sbodio J.I., Snowman A.M., Albacarys L., Hester L., Sedlak T.W., Paul B.D, & Snyder S.H. (2019). Bilirubin links heme metabolism to neuroprotection by scavenging superoxide. Cell chemical biology, 26(10), 1450-1460.e7.

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Osteogenic Differentiation Assay in Vitro

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Cells were cultured at 37°C with 5% CO₂ in osteogenic induction medium containing DMEM with 10% fetal bovine serum, β-glycerophosphate (10 mmol/l) and L-ascorbate (50 µg/ml), all purchased from Sigma-Aldrich (Merck KGaA), in 6-well plates at a density of 5×10⁴ cells/ml. When the cells reached 80% confluence, they were thrice rinsed with PBS and fixed with 95% ethanol for 10 min at the room temperature. ALP was stained with a BCIP/NBT kit (Beyotime Institute of Biotechnology, Haimen, China) and then observed using an inverted microscope (magnification, ×200). Following culture for 3 weeks, cells were fixed with 4% polyoxymethylene for 30 min at the room temperature, stained using alizarin red was performed at 37°C (Cyagen Biosciences Guangzhou, Inc., Guangzhou, China) and observed by an inverted microscope (magnification, ×40) to characterize the osteoblasts.

Liang D., Wang K.J., Tang Z.Q., Liu R.H., Zeng F., Cheng M.Y., Lian Q.S, & Wu H.K. (2018). Effects of nicotine on the metabolism and gene expression profile of Sprague-Dawley rat primary osteoblasts. Molecular Medicine Reports, 17(6), 8269-8281.

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HPLC Quantification of Vitamin C

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The concentration of vitamin C was evaluated according to a method described previously [24 (link)]. Samples were homogenized at 4 °C in 10 mL of 1% (0.01 g/mL) oxalic acid, and 5 mL of 1% oxalic acid were used to wash the residues twice. The combined extracts were then centrifuged at 7000 rpm for 10 min. The supernatant was filtered by a 0.45-μm cellulose acetate filter. The same system as the one used in the glucosinolate assay was employed to perform HPLC analysis, with a mobile phase of 0.1% oxalic acid at a flow rate of 1 mL/min. Absorbance values were collected at 243 nm. Standard (L-ascorbate; Sigma, St Louis, MO, USA) was prepared and used to identify and quantify the vitamin C. The results were expressed as mg/100 g fresh weight (FW).

Miao H., Lin J., Zeng W., Wang M., Yao L, & Wang Q. (2019). Main Health-Promoting Compounds Response to Long-Term Freezer Storage and Different Thawing Methods in Frozen Broccoli Florets. Foods, 8(9), 375.

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Preparing BHT and L-ascorbate for Cell Culture

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HU, butylated hydroxytoluene (BHT) and L-ascorbate were purchased from Sigma Aldrich (St. Louis, MO, USA) and prepared following the manufacturer's instructions. After complete solubilization, drugs were sterilized by filtration using a 0.22 μm polyethersulfone membrane (PES) (Jet Biofil, Guangzhou, China) for use in culturing assays.

Santana S.S., Pitanga T.N., de Santana J.M., Zanette D.L., Vieira J.D., Yahouédéhou S.C., Adanho C.S., Viana S.D., Luz N.F., Borges V.M, & Goncalves M.S. (2020). Hydroxyurea Scavenges Free Radicals and Induces the Expression of Antioxidant Genes in Human Cell Cultures Treated With Hemin. Frontiers in Immunology, 11, 1488.

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Osteoblast Differentiation using MC3T3-E1 Cells

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Osteoblast differentiation using the murine MC3T3-E1 cell line^{1 (link)} has been previously described.^{38 (link)} Briefly, MC3T3-E1 cells were differentiated to osteoblasts in αMEM supplemented with 10% FBS, penicillin, and streptomycin. Differentiation medium was supplemented with 50 μg of l-ascorbate and 10 mM β-glycerophosphate (Sigma Chemical Co., St. Louis, MO). NP1-MNP was added with medium changes twice per a week. Mineralization was visualized by staining with 40 mM Alizarin Red S (Sigma Chemical Co., St. Louis, MO) for 15 min. Excess stain was removed by washing with distilled water. Enzyme activity was measured in whole cell lysates using a kit from Sigma as described previously.^{38 (link)}

Ha S.W., Weitzmann M.N, & Beck GR J.r. (2014). Bioactive Silica Nanoparticles Promote Osteoblast Differentiation through Stimulation of Autophagy and Direct Association with LC3 and p62. ACS Nano, 8(6), 5898-5910.

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Antioxidant Reagents in Experiments

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L-Ascorbate, (L)-dehydroascorbate, menadione, and menadiol were purchased from Sigma-Aldrich (Weinheim, Germany). Other chemicals and kits were purchased from Santa Cruz Biotechnology Inc., Sigma-Aldrich, Wako, Abcam, Promega, and Cell Biolabs Inc.
All reagents, used in the experiments, were “analytical grade” or “HPLC grade”.

Bakalova R., Semkova S., Ivanova D., Zhelev Z., Miller T., Takeshima T., Shibata S., Lazarova D., Aoki I, & Higashi T. (2020). Selective Targeting of Cancerous Mitochondria and Suppression of Tumor Growth Using Redox-Active Treatment Adjuvant. Oxidative Medicine and Cellular Longevity, 2020, 6212935.

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