Eight-week-old male C57BL/6 mice were purchased from Vital River Laboratory Animal Technology, China. The study protocol was approved by the Committee on the Ethics of North China University of Science and Technology (LX2019033) and complied with the US National Institutes of Health Guide for the Care and Use of Laboratory Animals. The mice were randomly divided into three groups (n = 5) as follows: (1) the control group, subjected to a tracheal perfusion with 50 µL of 0.9% normal saline; (2) the silicosis group, subjected to a one-off non-invasive intratracheal instillation of silica suspension (100 mg/kg) (s5631; Sigma-Aldrich, St. Louis, MO, USA); and (3) the quercetin group, subjected to an intraperitoneal injection of quercetin (100 mg/kg) (Q4951, Sigma–Aldrich, Shanghai, China) every day for 28 days at the time of silica suspension (100 mg/kg) via a one-off non-invasive intratracheal instillation. The mice were sacrificed and part of the lung tissue was dehydrated and embedded in paraffin, while the remaining lung tissue was stored at −80 °C.
Q4951
Manufactured by Merck Group
Sourced in United States
The Q4951 is a laboratory equipment product manufactured by Merck Group. It serves as a core function in various scientific and research applications, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
Quercetin, by Merck Group (6 mentions)
Dasatinib, by Merck Group (3 mentions)
S5631, by Merck Group (1 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Valproic acid, by Merck Group (1 mentions)
Suberoylanilide hydroxamic acid saha, by Merck Group (1 mentions)
A2385, by Merck Group (1 mentions)
P4543, by Merck Group (1 mentions)
5 azacytidine, by Merck Group (1 mentions)
Sodium butyrate, by Merck Group (1 mentions)
C9625, by Merck Group (1 mentions)
Ftp 20 30, by Instech (1 mentions)
Peg400, by Merck Group (1 mentions)
Gm08400, by Coriell Institute for Medical Research (1 mentions)
Sml2589, by Merck Group (1 mentions)
Insulin transferrin selenium, by Thermo Fisher Scientific (1 mentions)
Gelatin based coating solution, by Cell Biologics (1 mentions)
Iron 3 chloride fecl3, by Merck Group (1 mentions)
Dextran, by Merck Group (1 mentions)
Iron 2 chloride fecl2, by Merck Group (1 mentions)
16 protocols using q4951
1
Silica-induced Pulmonary Fibrosis in Mice
2
Epigenetic Modifiers Protocol
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Six epigenetic modifiers were opted based on literature review viz., two DNMT inhibitors 5-azacytidine (A2385, Sigma-Aldrich) and hydralazine hydrochloride (H1753, Sigma-Aldrich), a sirtuins activator [quercetin (Q4951, Sigma-Aldrich)] and three HDAC inhibitors Suberoylanilide Hydroxamic Acid (SAHA) (390585, Sigma-Aldrich), sodium butyrate (303410, Sigma-Aldrich) and valproic acid (P4543, Sigma-Aldrich)] (de la Cruz et al., 2012 (link); González-Menéndez et al., 2016 (link)).
3
Quercetin, Carnosine, and Ozone in Semen
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Quercetin (Sigma-Aldrich – Q4951) was diluted in dimethyl sulfoxide (DMSO) to a final concentration of 0.1% after incorporation into semen. This concentration corresponds to the maximum non-harmful concentration of DMSO in the sperm (Meamar et al., 2012 (link)). Solid carnosine (Sigma-Aldrich – C9625), previously diluted in deionized water, was used.
Ozone was obtained using a portable ozone generator model O&L 1.5 (Ozone & Life™), programmed to release a flow of medicinal ozone (O3) sufficient to produce the concentrations indicated for the experimental groups. A 5 mL sterile syringe was attached to the device before starting the process, where 2.5 mL of O3 gas was stored immediately before contacting the semen. For O3 incorporation, 2.5 mL of semen was mixed with the gas inside the syringe for 30 s.
Ozone was obtained using a portable ozone generator model O&L 1.5 (Ozone & Life™), programmed to release a flow of medicinal ozone (O3) sufficient to produce the concentrations indicated for the experimental groups. A 5 mL sterile syringe was attached to the device before starting the process, where 2.5 mL of O3 gas was stored immediately before contacting the semen. For O3 incorporation, 2.5 mL of semen was mixed with the gas inside the syringe for 30 s.
4
Senolytic Cocktail Administration in Mice
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Mice were administered a senolytic cocktail containing 5 mg/kg dasatinib (D‐3307, LC Labs) and 50 mg/kg quercetin (Q4951, Sigma‐Aldrich) as described by Xu et al. (Xu et al., 2018 (link)). Briefly, 7.5 mg of dasatinib and 75 mg of quercetin were dissolved in 5 ml of 10% polyethylene glycol 400 (PEG 400; 202398, Sigma‐Aldrich). We chose this volume of PEG 400 because it allowed us to gavage a 30–45 gram mouse with 100–150 μl of D+Q, which is less than the approximate stomach volume of 400 μl in adult mice (McConnell et al., 2008 (link)). Mice were gavaged bi‐weekly for 4 months (Figure 1a ) with D+Q (n = 20 young and n = 19 old) or vehicle (n = 20 young and n = 19 old; 10% PEG 400) using 20‐gauge disposable polypropylene feeding tubes (FTP‐20‐30, Instech, Plymouth Meeting, PA). These mice were then divided into 3 groups of n = 6–7/group.
5
Fibroblast Modeling of Myotonic Dystrophy
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DM1-A, DM2, and unaffected control-A fibroblasts were previously obtained from skin biopsies under a University of Florida-approved Institutional Review Board (IRB) protocol with the informed consent from all subjects and have been described previously (22 (link)). DM1-B (GM03987) and unaffected control-B (GM08400) fibroblasts were obtained from Coriell Institute. Approximately 1×105 cells were seeded per well of 12 well culture plates and cultured to ~100 % confluence in DMEM or MEM media containing 15% FBS and 1% antimycotic/antibacterial under standard conditions of 37°C and 5% CO2. Once confluency was reached, cells were washed with PBS and fresh growing media containing quercetin (Sigma-Aldrich Q4951), dasatinib (Sigma-Aldrich SML2589) or A1155463 (Sigma-Aldrich SML3162) dissolved in DMSO was added at the indicated concentrations for 24 hours.
6
Isolation and Senescence Depletion of Type II Alveolar Epithelial Cells
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AECs were isolated by magnetic associated cell sorting (MACS) by negative selection of CD45+ (Miltenyi, cat#130-052-301, dilution 10 µL per 10 million cells) and CD31+ (Miltenyi, cat#130-097-41, dilution 10 µL per 10 million cells) cells and followed by positive selection of CD326+ cells (Miltenyi, cat# 130-118-075, clone caa7-9G8, dilution 5 µL per 10 million cells). This method typically yields approximately 85% purity of prosurfactant-C+ cells as measured by flow cytometry. Isolated type II AECs were cultured with DMEM/F12 medium containing 10% FBS, 1.25 g BSA, 100 U/mL penicillin/streptomycin, and 1x Insulin-Transferrin-Selenium (Gibco, 41400045). Type II AECs were seeded in tissue culture plates coated with gelatin-based coating solution (Cell Biologics, 6950).
To deplete senescent cells, type II AECs were cultured with 20 nM dasatinib (Sigma, SML2589) and 5 µM quercetin (Sigma, Q4951) or 200 nM dasatinib and 50 µM quercetin for 48 h.
To deplete senescent cells, type II AECs were cultured with 20 nM dasatinib (Sigma, SML2589) and 5 µM quercetin (Sigma, Q4951) or 200 nM dasatinib and 50 µM quercetin for 48 h.
7
Spectrophotometric Quantification of Honey Flavonoids
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The total flavonoid (TF) content was determined spectrophotometrically. Briefly 1 mL of honey extract or a standard solution of quercetin (Sigma Q4951) (10, 50, 100, 150, 200, and 250 μg/mL) in distilled water was added to a 10 mL volumetric flask containing 4 mL of double distilled water, 300 μL of NaNO2 (5%, v/v), and 300 μL of 10% AlCl3. The solution was allowed to stand at room temperature in the dark for 30 minutes and the absorbance was read at 430 nm. The TF content was determined using the standard curve of quercetin (μg/mL) and was expressed as μg of quercetin equivalents in 1 g of extract.
8
Synthesis and Characterization of Qu-Dextran Nanoparticles
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All chemical material required for QNPs synthesis including Qu (≥95% purity, Q4951, molecular weight: 302.2 g.mol−1), dextran (molecular weight: 10,000 Da), Iron (III) chloride (FeCl3), and iron (II) chloride (FeCl2) were obtained from Sigma-Aldrich Co. (St Louis, MO, USA) or EMD Millipore Co. (Billerica, MA, USA). The materials required for cell culture including phosphate buffer saline (PBS), RPMI 1640, and others were purchased from Bioidea Co. (Tehran, Iran).
9
Extraction and Analysis of Phytochemicals
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Solvents used for the extraction and analysis of phytochemicals (methanol, isopropanol, acetonitrile, water and formic acid) were LC-MS grade (67-56-1, 67-63-0, 75-05-8, 7732-18-5, 85178, respectively, Sigma-Aldrich, (St. Louis, MO, USA). Authentic standards for mangiferin, (+)-catechin, quercetin-3-D-galactoside, quercetin, gallic acid, (-)-epicatechin and quercetin-3-glucoside (M3547, 43412, 83388, Q4951, G7384, 1753, 16654, respectively, Sigma-Aldrich, St. Louis, MO, USA) were used. Kaempferol-3-O-glucoside (90242), 4-hydroxy-benzoic acid (99-96-7), caffeic acid (6034 S), 4-coumaric acid (6031 A), ferulic acid (6077 A), quercetin 3, 4-di-O-glucoside (1347 S), quercetin 3-D-O-galactoside (1027 S) standards were purchased from Extrasynthese (https://www.extrasynthese.com/ ). All reagents used for proteomic analysis were purchased from Sigma-Aldrich (St. Louis, MO, USA), except as otherwise specified in the corresponding section.
10
Comparative Evaluation of Antioxidant Compounds
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The cells were treated with different concentrations of resveratrol (R5010, Sigma-Aldrich), Epigallocatechin gallate (EGCG, E4143, Sigma-Aldrich), and quercetin (Q4951, Sigma-Aldrich) dissolved in ethanol (EtOH). The highest concentration of EtOH (0.18%, 0.07%, and 0.05% for quercetin, EGCG, and resveratrol, respectively) applied to the cells during treatment was used as a control in order to eliminate EtOH effects from our data. Furthermore, the data for quercetin were normalized to an EtOH curve, with the same concentrations as those used during the substance treatment, to further reduce the effect of the solvent.
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