Cell culture materials were obtained from PAA laboratories (Pasching, Austria). Oligomycin A, histamine, 2,5-di-t-butyl-1,4-benzohydroquinone (BHQ), and ethylene glycol tetraacetic acid (EGTA) were purchased from Sigma Aldrich (Vienna, Austria), thapsigargin, piceatannol, and resveratrol from Abcam (London, UK). Prior to experiments, cells were washed and maintained for 20 minutes in a HEPES-buffered solution containing 138 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 1 mM HEPES, 2.6 mM NaHCO3, 0.44 mM KH2PO4, 0.34 mM Na2HPO4, 1 mM D-glucose, 0.1% vitamins, 0.2% essential amino acids, and 1% penicillin-streptomycin, the pH of which was adjusted to 7.4 with NaOH or HCl. During the experiments cells were perfused with a Ca2+-containing buffer, which consisted of 145 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 10 mM D-glucose and 10 mM HEPES, pH adjusted to 7.4, or with a Ca2+-free buffer, in which CaCl2 was replaced by 1 mM EGTA.
Resveratrol
Manufactured by Abcam
Sourced in United Kingdom
Resveratrol is a natural polyphenol compound found in various plant species, including grapes, berries, and peanuts. It is a potent antioxidant and has been studied for its potential health benefits. Resveratrol is often used in research applications to investigate its effects on cellular processes and physiological functions.
Automatically generated - may contain errors
Lab products found in correlation
Thapsigargin, by Abcam (2 mentions)
Oligomycin a, by Merck Group (2 mentions)
Histamine, by Merck Group (2 mentions)
2 5 di t butyl 1 4 benzohydroquinone, by Merck Group (2 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (2 mentions)
Piceatannol, by Abcam (2 mentions)
Dapi d9542, by Merck Group (1 mentions)
U87mg, by American Type Culture Collection (1 mentions)
Mitotracker red, by Thermo Fisher Scientific (1 mentions)
β actin, by Proteintech (1 mentions)
Bafilomycin a1 ab120497, by Abcam (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
As2o3, by Merck Group (1 mentions)
Spironolactone, by Selleck Chemicals (1 mentions)
Veliparib, by Selleck Chemicals (1 mentions)
Trypsin edta, by Biosera (1 mentions)
Abt 888, by Selleck Chemicals (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Biosera (1 mentions)
Fluoxetine hydrochloride, by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
8 protocols using resveratrol
1
Calcium Signaling Cell Assay Protocol
2
Resveratrol Therapy for Surgical Mice
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In the fourth experiment, some mice received daily intraperitoneal injection of 50 mg/kg resveratrol (Abcam, Cambridge, UK; dissolved in 25% DMSO to a final concentration of 10 mg/mL) based on a previous study [30 (link)] and some mice received intraperitoneal injection of the DMSO solvent (sham solution). This injection was started 2 h after the surgery and was for 7 days.
3
Autophagy Regulation by Sirtuins
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U87MG and T98G cells were purchased from ATCC. Cells were cultured in Dulbecco’s Modified Eagle Medium with 10% fetal bovine serum and incubated with 5% CO2. All candidate compounds tested in this study were purchased from Life Chemicals Inc (Kyiv, Ukraine). MTT (M2128), 3-MA (M9281), and DAPI (D9542) were purchased from Sigma-Aldrich (St. Louis, MO, USA). MitoTracker Red was purchased from Thermo Fisher Scientific Inc. (NY, USA). Bafilomycin A1 (ab120497) and Resveratrol (ab120726) were purchased from Abcam (Cambridge, UK). Antibodies used in this study were as follow: SIRT1 (2496, CST, MA, USA), Acetyl-Histone H3 (9649, CST), Histone H3 (4499, CST), p53 (2527, CST), Acetyl-p53 (2525, CST), AMPK (5831, CST), p-AMPKthr172 (2535, CST), ULK1 (8054, CST, MA, USA), p-ULK1ser317 (12753, CST), p-ULK1ser555 (12753, CST), mAtg13 (13273, CST), Atg101 (13492, CST), FIP200 (12436, CST), mTOR (2983, CST), p-mTORser2448 (12436, CST), Beclin-1 (3495, CST), LC3B (3868, CST), SQSTM1/p62 (5114, CST), BNIP3 (44060, CST), PINK1 (6946, CST), Parkin (4211, CST), VDAC1 (4661, CST), ATG5 (12594, CST), ATG12 (4180, CST), ATG16L (8089, CST), 14-3-3γ (5522, CST), catalase (12980, CST), Profilin-1 (3237, CST), HSP90 (4877, CST), β-actin (66009-1-Ig, Proteintech, IL, USA).
4
UVB-Induced Cellular Stress Assays
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Cells were harvested with trypsin-EDTA (Biosera, Budapest, Hungary) and then seeded in six-well plates for the hypoxanthine phosphoribosyltransferase (HPRT) gene mutation assay or 24-well plates for all other experiments. At ~80% confluence, cells were pretreated with 25 μM ABT-888 (PARP1 inhibitor, veliparib, Selleckchem, Houston, TX, USA), 10–50 μM resveratrol (Abcam, Cambridge, UK), 5–25 μM spironolactone (Selleckchem, Houston, TX, USA), or 0.5–4 μg/mL As2O3 (Sigma-Aldrich, St. Louis, MO, USA) solution. In the case of veliparib treatment, we chose the concentration that caused marked inhibition of PARP1 protein, according to our previous [29 (link)] and current experiments (Figure S9 ). For the other chemicals, we identified three different concentrations due to their more complex and not fully understood mode of action—based on prior published data [26 (link),27 (link),30 (link),31 (link),32 (link)]. As2O3 was dissolved in 1 M NaOH (Sigma-Aldrich, St. Louis, MO, USA) and diluted in Dulbecco’s phosphate-buffered saline (DPBS; Biosera, Budapest, Hungary). Other chemicals were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA). Pretreated cells were incubated for 120 min at 37 °C before UVB irradiation.
5
Calcium Signaling Cell Assay Protocol
6
Neuroprotective Effects of Resveratrol and 3-MA in SAH
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One hundred and forty-one Sprague-Dawley rats were distributed randomly into five groups: the sham group (n=30), the SAH group (n=60), the SAH+Resveratrol (n=30), and SAH+3-methyladenine (3-MA) (n=21). Animals in the sham group received 0.9% saline (intraperitoneal injection) after the sham operation. The SAH group received an equal amount of DMSO solution (0.1% dimethylsulfoxide in 0.9% saline). After SAH, the SAH+Resveratrol group received 60 mg/kg Resveratrol (intraperitoneal injection) 13 (link) and the SAH+3-MA group received 60 mM 3-MA (intracerebroventricular administration) 14 (link). Resveratrol and 3-MA were purchased from Abcam (ab120726; Cambridge, UK) and Cayman (13242; Ann Arbor, Michigan, USA), respectively.
7
Lipopolysaccharide-induced Oxidative Stress
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Lipopolysaccharide (LPS) (Escherichia coli serotype O111:B4), fluoxetine hydrochloride, 2-thiobarbituric acid (TBA), sodium dihydrogen phosphate anhydrous, disodium hydrogen phosphate anhydrous and trichloroacetic acid were purchased from Sigma-Aldrich (Sigma-Aldrich Co. LLC (St Louis, MO, USA). Resveratrol and sirtinol were procured from Abcam (Abcam plc, Cambridge, UK). All other chemicals used in this study were of analytical grade.
8
Resveratrol effects on immortalized mouse HSCs
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The immortalized mouse HSC line JS1 was kindly provided by Professor Scott L. Friedman (Mt. Sinai School of Medicine, USA). JS1 cells were cultured in DMEM (Gibco; Thermo Fisher Scienti c, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scienti c, Inc.) and 1% penicillin/streptomycin at 5% CO 2 and 37°C. The medium was replaced with media supplemented with 0.5% FBS for 12 h prior to treatment. Different concentrations of resveratrol (cat. no. R5010; Sigma) was added to the medium and an incubated for different durations, according to the experimental design. JS1 cells were pretreated with 5 mM 3-methyladenine (3-MA; cat. no. M9281; Sigma-Aldrich; Merck KGaA), 30 µM chloroquine (CQ) diphosphate salt (cat. no. C6628; Sigma-Aldrich; Merck KGaA) or 20 µM Z-VAD-FMK (cat. no. C1202; Beyotime Institute of Biotechnology) for 2 h and 10 µM EX527 (cat. no. ab141506; Abcam) or 10 µM SP600125 (cat. no. ab120065; Abcam) for 1 h, followed by treatment with or without 50 µM resveratrol for 24 h. Each in vitro experiment was repeated three times.
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