Detection of cell viability was via the cell counting kit (CCK)-8 method. Seeding of MC3T3-E1 cells was into 96-well plates at a density of 5 × 103 cells/well, and incubation for 24 h, 48 h, and 72 h under the following three conditions: DMEM (Control), DMEM + 30 μM H2O2 (H2O2), DMEM + 1 µM PBC + 30 μM H2O2 (1 µM PBC + H2O2) and DMEM + 10 µM PBC + 30 μM H2O2 (10 µM PBC + H2O2). The concentration of probucol (Sigma, St. Louis, MO, USA) was determined based on the published in vitro studies on probucol (Li et al. 2021a (link)). Then, 10 mL CCK-8 reagent was in each well. Measurement of the absorbance was at 450 nm with a microplate reader. Performance of cytotoxicity tests was evaluated with lactate dehydrogenase (LDH) detection kits (Nanjing Jiancheng, Nanjing, China) in line with the manufacturer’s agreement.
Probucol
Manufactured by Merck Group
Sourced in United States, United Kingdom, Australia
Probucol is a laboratory equipment product manufactured by Merck Group. It is a chemical compound used for research and analytical purposes. Probucol has specific chemical and physical properties that make it suitable for various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Streptozotocin (stz), by Merck Group (2 mentions)
Ursodeoxycholic acid, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Low viscosity sodium alginate, by Merck Group (2 mentions)
Ldh detection kit, by Nanjing Jiancheng (1 mentions)
Lc ms grade methanol, by Thermo Fisher Scientific (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Fumaric acid, by Merck Group (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
Paclitaxel, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Rapamycin, by Thermo Fisher Scientific (1 mentions)
Glucose 6 phosphate (g6p), by Merck Group (1 mentions)
Choline chloride, by Merck Group (1 mentions)
Ibuprofen, by Merck Group (1 mentions)
Trifluoroacetic acid tfa, by Thermo Fisher Scientific (1 mentions)
Superfrost plus, by Thermo Fisher Scientific (1 mentions)
Dopamine hydrochloride, by Merck Group (1 mentions)
Raffinose, by Merck Group (1 mentions)
Glutamic acid, by Merck Group (1 mentions)
23 protocols using probucol
1
Cell Viability Assay of MC3T3-E1 Cells
2
Probucol Neuroprotection in Huntington's Disease
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Probucol (Sigma, St. Louis, MO, USA; 30 mg/kg/day) was diluted in carboxymethylcellulose (CMC), 1 mg/mL. The Probucol dose was chosen based on previous studies that reported neuroprotective effects of this compound [48 (link)–50 (link)]. YAC128 and wild-type (WT) mice were randomly divided into four experimental groups (with equal numbers of males and females included in each group, 10 mice/group): 1 (WT vehicle), 2 (YAC128 vehicle), 3 (WT Probucol), and 4 (YAC128 Probucol). Probucol-treated WT and YAC128 mice received Probucol in drinking water for 5 months (from 1 to 6 months of age). Vehicle-treated WT and YAC128 mice received vehicle (CMC) in drinking water during the same period of time (Figure 1 ).
3
MALDI-TOF Lipid Profiling Protocol
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Methanol (LC-MS grade), Fisher Scientific (Leicestershire, UK); purified water, ELGA Purelab Option (Marlow, UK); Lipid standard phosphatidylcholine (PC) 16:0/18:1, Avanti Polar Lipids (Delfzyl, The Netherlands); Trifluoroacetic acid (TFA), Acros Organics (Loughborough, UK); MALDI matrices and analytes 2,6-dihydroxyactophenone (2,6-DHAP), histidine, glutamic acid, amiodarone, insulin, probucol, choline chloride, fumaric acid, cysteine, ketoglutaric acid, glutamic acid, dopamine hydrochloride, l-histidine, tryptophan, ibuprofen, palmitic acid, glucose 6 phosphate, glutathione, clozapine, glycochenodeoxycholate, raffinose, probucol, amiodarone, co-enzyme Q10, ubiquitin (from bovine erythrocytes) were purchased from Sigma Aldrich (Dorset, UK) at 98% purity or above. Paclitaxel and rapamycin from Alfa Aesar (Lancashire, UK). Superfrost Plus (Thermo Scientific, Waltham, MA, USA) glass slides were used for all experiments.
4
hERG-HEK293 Cells Exposure to Probucol and CCh
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5
Antioxidant and Anti-Inflammatory Assay
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Probucol, anti-β-actin antibody, hexadecyltrimethylammonium bromide, o-dianisidine dihydrochloride, and hematoxylin and eosin were purchased from Sigma Chemical Company (St. Louis, MO, USA). Protein assay kit was obtained from Bio-Rad Laboratories (Hercules, CA, USA). The anti-E-selectin antibody, horseradish peroxidase-linked secondary antibody, and enhanced chemiluminescence reagent were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Malondialdehyde assay kit was obtained from Nanjing Jiancheng Institute of Bioengineering (Nanjing City, China). All other reagents were of the highest grade commercially available.
6
Silane-functionalized Probucol Synthesis
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Tetraethyl orthosilicate (TEOS) and 3-aminopropyl triethoxysilane (APTES) were purchased from Sigma Aldrich (Sydney, Australia) as reagent grade, and used without further purification. Throughout the experiments Milli-Q water was used. Probucol was purchased from Sigma Aldrich (Sydney, Australia) and used without further purification.
7
Probucol Improves Diabetic Outcomes
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Thirty 12-week-old Sprague–Dawley male rats were obtained from the Animal Center of Shandong University (Jinan, China). The present experimental design was approved by the Animal Care and Use Committee of Shandong University (Jinan, China). DM was induced by a single intraperitoneal injection of 60 mg kg−1 STZ (Sigma-Aldrich, St. Louis, MO, USA) in 20 Sprague–Dawley rats. Ten rats were given vehicle only (0.1 mol l−1 citrate–phosphate buffer, pH 4.5) and used as a sham group. Animals with blood glucose levels consistently greater than 16.7 mmol l−1 were considered diabetic after 72 h. Twenty diabetic rats were established successfully, that is, 20 rats were all diabetic rats. After 12 weeks, the diabetic animals were divided randomly into two treatment groups: the diabetic control group (DM group, n = 10) and the experimental group (probucol group, n = 10). The experimental group received a daily gavage of probucol (Sigma-Aldrich) at a dose of 300 mg kg−1 for 12 weeks. The diabetic control group received physiological saline only. The body weights and blood glucose levels of all rats were recorded weekly.
8
MTT Assay for Cytotoxicity Evaluation
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Twenty-four hours after plating, cells were exposed to drugs to be tested: cisplatin (Accord, Italy), and/or fendiline hydrochloride (Sigma Aldrich, cat. Number F7265) and/or 1 µM sulfasalazine (Sigma Aldrich, cat. Number S0883) and/or 5µM probucol (Sigma Aldrich, cat. Number P9672), at 37 °C. At the end of the treatment, the medium was removed and cells were incubated for 1 h with 3-(4,5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT) solution (2 mg/mL in PBS) (Sigma-Aldrich). In order to dissolve formazan crystals, after removing MTT 500 μL of 100% ethanol were added to each well, and absorbance was determined at 570 nm, using a reference filter at 670 nm. Cytotoxicity was expressed as percentage of viable cells compared to untreated cells.
9
Multiparametric Cell Viability Assay
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10
Curcumin Modulates Cell Viability and Apoptosis
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The reagents included DMEM/F12 high glucose (Hyclone, Utah, USA), penicillin (Hyclone, Utah, USA), streptomycin (Hyclone, Utah, USA), curcumin (Bellancom, Beijing, China), trypsin (Google Biotechnology, Wuhan, China), collagenase-II, bovine serum albumin (BSA), probucol (Sigma-Aldrich, St. Louis, MO, USA), KeyFluor488 Click-iT EdU kits, DAPI, (KeyGEN BioTECH, Nanjing, China), AnnexV-PI kits (BD, USA), Counting Kit-8 (CCK-8) reagents, goat serum (Beyotime Institute of Biotechnology, Shanghai, China), TNF-α (Peprotech, Inc., Suzhou, China), Caspase-3, Bcl-2, Lc3, Bax, PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR, GADPH, Berclin-1, COL-II, P62, FITC, Cy3, MMP-3, JC-1 assay kits (Abcam, USA), TRIzol reagents (Invitrogen, Thermo Fisher Scientific, Inc. USA), a RevertAid First Strand cDNA Synthesis kit (Fermentas; Thermo Fisher Scientific, Inc. USA), MMP detection kits (Solarbio Science & Technology, Beijing, China), and chemiluminescent luminol reagent (Santa Cruz Biotechnology, Texas, USA).
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