T. Fructus was purchased from a local traditional Chinese medicine market in Henan province, China. Sephadex G-100 column was obtained from GE Healthcare (Beijing, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), and ascorbic acid (VC), were obtained from Sigma (St. Louis, MO, USA). Cellulase (10,000 u/g) was purchased from Jinsui Biological Technology Co. Ltd. (Shanghai, China). All other analytical-grade chemicals were obtained from Nanjing Reagent Co. Ltd. (Nanjing, China).
Ascorbic acid vc
Manufactured by Merck Group
Sourced in United States
Ascorbic acid, also known as Vitamin C, is a white crystalline solid that is commonly used in various laboratory applications. It is a water-soluble vitamin that functions as an essential nutrient and antioxidant. Ascorbic acid is widely used in research and analysis due to its chemical properties and versatility.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Ripa cell lysis buffer, by Thermo Fisher Scientific (2 mentions)
Streptavidin agarose bead, by Thermo Fisher Scientific (2 mentions)
Biotin nem, by Thermo Fisher Scientific (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Sephadex g 100 column, by GE Healthcare (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Antibodies for flow cytometry, by BioLegend (1 mentions)
Sodium selenite na2seo3, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Bca assay kit, by Beyotime (1 mentions)
Trypsin, by Merck Group (1 mentions)
Pepsin, by Merck Group (1 mentions)
Acarbose, by Merck Group (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Na2seo3, by Merck Group (1 mentions)
Parp antibody, by Cell Signaling Technology (1 mentions)
Supersignal chemiluminescent detection system, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
16 protocols using ascorbic acid vc
1
Purification and Antioxidant Evaluation of Fructus
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T. Fructus was purchased from a local traditional Chinese medicine market in Henan province, China. Sephadex G-100 column was obtained from GE Healthcare (Beijing, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), and ascorbic acid (VC), were obtained from Sigma (St. Louis, MO, USA). Cellulase (10,000 u/g) was purchased from Jinsui Biological Technology Co. Ltd. (Shanghai, China). All other analytical-grade chemicals were obtained from Nanjing Reagent Co. Ltd. (Nanjing, China).
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T. Fructus was purchased from a local traditional Chinese medicine market in Henan province, China. Sephadex G-100 column was obtained from GE Healthcare (Beijing, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), and ascorbic acid (VC), were obtained from Sigma (St. Louis, MO, USA). Cellulase (10,000 u/g) was purchased from Jinsui Biological Technology Co. Ltd. (Shanghai, China). All other analytical-grade chemicals were obtained from Nanjing Reagent Co. Ltd. (Nanjing, China).
2
Antioxidant Evaluation in Cell Cultures
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GLSO was purchased from certain commercial company. Sodium selenite (Na2SeO3) and ascorbic acid (VC) were purchased from Sigma Aldrich (USA). PI, JC-1, and glutathione peroxidase (GSH-Px), SOD, malondialdehyde (MDA), CCK8, and BCA assay kits were obtained from Beyotime Biotechnology (China). Dulbecco’s modified eagle medium (DMEM), 1640 medium, penicillin–streptomycin, and fetal bovine serum (FBS) were purchased from Gibco (USA). Antibodies for flow cytometry were obtained from Biolegend (USA), and antibodies for Western blot (WB) and ELISA were obtained from Cell Signaling Technology (USA).
3
Antioxidant Activity Assay
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1,1-Diphenyl-2-picrylhydrazyl (DPPH•), pepsin (3640 U mg−1 protein), trypsin (3640 U mg−1 protein), acarbose, and ascorbic acid (VC) were purchased from Sigma (St. Louis, MO, USA). All other chemicals used in the experiments were of analytical reagent grade from Sangong Biotech Co., Ltd. (Shanghai, China).
4
EV71 Infection in Neuroblastoma Cells
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A human
neuroblastoma cell line (SK-N-SH)
was bought from Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences. Dulbecco’s modified eagle medium (DMEM)
and fetal bovine serum (FBS) were procured from Gibco. EV71 virus
was isolated and stored in Guangzhou Women and Children’s Medical
Center (GenBank Accession FJ360545.1). Na2SeO3 and ascorbic acid (VC) from Sigma were used in the study. The one-step
qPCR kit was acquired from Takara. SiRNAs and primers were designed
and synthesized by Sangon Biotech, China. The VP1 monoclonal antibody
was bought from Abnova. Bax and β-actin monoclonal antibodies
were supplied by Cell Signaling Technology.
neuroblastoma cell line (SK-N-SH)
was bought from Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences. Dulbecco’s modified eagle medium (DMEM)
and fetal bovine serum (FBS) were procured from Gibco. EV71 virus
was isolated and stored in Guangzhou Women and Children’s Medical
Center (GenBank Accession FJ360545.1). Na2SeO3 and ascorbic acid (VC) from Sigma were used in the study. The one-step
qPCR kit was acquired from Takara. SiRNAs and primers were designed
and synthesized by Sangon Biotech, China. The VP1 monoclonal antibody
was bought from Abnova. Bax and β-actin monoclonal antibodies
were supplied by Cell Signaling Technology.
5
Quantifying S-nitrosated Proteome by Biotin-Switch
6
DPPH Radical Scavenging Assay for EPS
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The scavenging activity for DPPH was assayed as described in the study by Niknezhad et al. [36 (link)]. The reaction mixture contained 50 μL of EPSt at different concentrations (0.1, 0.25, 1.0, 2.5, 5.0, 7.5 and 10 mg/mL) and 100 μL of DPPH (100 μM DPPH–ethanolic solution) (Sigma Chemical, St Louis, MO, USA). The mixtures were shaken vigorously and incubated in the dark at 25 °C. After 30 min, the absorbance was recorded at OD525nm. Ascorbic acid (Vc) (Sigma Chemical, St Louis, MO, USA) was used as positive control.
The percentage of radical scavenging activity for DPPH was calculated according to Formula (1):
where
A1 = OD525 nm of reaction mixture.
A2 = OD525 nm of reaction mixture without DPPH.
A0 = OD525 nm of the reaction mixture with DPPH but without EPSt.
The percentage of radical scavenging activity for DPPH was calculated according to Formula (1):
where
A1 = OD525 nm of reaction mixture.
A2 = OD525 nm of reaction mixture without DPPH.
A0 = OD525 nm of the reaction mixture with DPPH but without EPSt.
7
Osteogenic and Adipogenic Differentiation of Fourth-Generation Cells
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Each group’s fourth-generation cells were trypsinized and seeded in a six-well plate at a density of 3 × 103 cells/well, and 2 mL of α-MEM medium containing 5% fetal bovine serum (FBS) was added to each well and placed at a constant temperature in a cell incubator. After 24 h, each well was replaced with osteogenic induction medium (α-MEM medium with 5% FBS, dexamethasone 10 mol/L, ascorbic acid Vc 50 µg/mL, and β-glycerophosphate sodium 10 mmol/L; Sigma-Aldrich) and adipogenic induction medium (α-MEM medium with 5% FBS, dexamethasone 0.25 µmol/L, indomethacin 100 mmol/L, IBMY 0.5 mmol/L, and insulin 10 mg/L; Sigma-Aldrich) to continue culturing. The medium was changed every three days for three to four weeks during induction. Each group was stained with alizarin red and oil red O (Sigma-Aldrich). When staining, the cell induction medium was aspirated, washed three times with PBS, and filtered with 4% paraformaldehyde (Beyotime, China) for 20 min. Then, the medium was discarded and washed three times with PBS. After adding alizarin red or oil and drying red O, cells were incubated at 37 ℃ for 2 h. After discarding the dye, the sample was rinsed three times with PBS and photographed under an inverted microscope.
8
Biphasic Calcium Phosphate Ceramic and Collagen I
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Fresh biphasic calcium phosphate ceramic (BCP, HA : β-TCP = 2 : 8, ϕ10 × 2 mm, ϕ4 × 3 mm) and type I collagen (Col I, extracted from calf skin) were provided by National Engineering Research Center in Biomaterials, Sichuan University. Phosphate-buffered saline (PBS) and α-minimum essential medium (α-MEM) were purchased from Thermo Fisher Scientific Corporation (USA). Fetal bovine serum (FBS, Gibco, Australia origin) was purchased from Life Technologies Corporation (USA). Ascorbic acid (Vc) was purchased from Sigma-Aldrich.
9
Biotin-Switch Assay for S-Nitrosation
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This assay is modified from the “biotin-switch” assay [33 (link)–35 (link)]. All following steps were performed in the dark. Treated HEKn cells were harvested in RIPA cell lysis buffer (Thermo Scientific), sonicated, and centrifuged at 13,500 rpm for 15 min at 4°C to remove cellular debris. 10 mM N-ethylmaleimide (NEM; Sigma-Aldrich) was added and incubated for 1 h at room temperature to block all free Cys residues. Excess NEM in samples was removed by ice-cold acetone precipitation repeated three times for 20 min each. 1 mM ascorbic acid (Vc, Sigma-Aldrich) and 500 μM biotin-NEM (Thermo Scientific Pierce) were then added and incubated at room temperature for 1 h in order to reduce S-nitrosated Cys on proteins and label the residues. Labeled S-nitrosated proteome was purified using streptavidin agarose beads (Thermo Scientific Pierce) for further immunoblotting analysis.
10
Thermoascus aurantiacus Heterologous Protein Expression
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Thermoascus aurantiacus strain CGMCC3.17992 was isolated from horse manure in China according to our previous method (Li et al., 2003 (link)) and conserved in the China Microbial Strain Collection (Beijing, China). The plasmids pPICZαA and P. pastoris GS115 were purchased from Invitrogen. Avicel PH-101, ascorbic acid (Vc), glucose, gluconic acid, galactose, and sorbitol were purchased from Sigma. The culture medium mainly used low-salt LB medium, YPDS medium, BMGY, and BMMY medium.
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