Human uterine leiomyoma (UtLM) cells (GM10964; Coriell Institute for Medical Research, Camden, NJ, USA) and uterine smooth muscle cells (UtSMC) (Clonetics Corporation, San Diego, CA, USA) were kept in a standard tissue culture incubator at 37°C, with 95% humidity and 5% carbon dioxide. The UtLM cells were routinely cultured in Minimum Essential Medium (MEM) Eagle (Sigma Chemical Company, St. Louis, MO, USA) as previously described [28 (link)]. The UtSMC were cultured in Smooth Muscle Cell Growth Media System (SmGM-2 BulletKit®) (Lonza, Basel, Switzerland) as previously reported [28 (link)]. The media were changed 24 h prior to genistein treatment to DMEM/F-12 (Sigma) phenol red free with charcoal/dextran treated fetal bovine serum (FBS) (Hyclone Laboratories, Logan, UT, USA) for both cell types. The cells were then treated with genistein (4’, 5, 7-Trihydroxyisoflavone; ≥ 98% purity by HPLC) (Sigma) every two days until day seven. genistein was reconstituted using DMSO (≥ 99.7 % purity by HPLC) (Sigma) before it was diluted into the media.
Genistein
Manufactured by Merck Group
Sourced in United States, Germany, China, France, United Kingdom, Sao Tome and Principe, Italy, Australia, Poland
Genistein is a lab equipment product from Merck Group. It is a naturally occurring isoflavone compound found in various plants. Genistein can be used as a research tool in various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Daidzein, by Merck Group (94 mentions)
Quercetin, by Merck Group (44 mentions)
Genistin, by Merck Group (39 mentions)
Chlorpromazine, by Merck Group (39 mentions)
Daidzin, by Merck Group (33 mentions)
Apigenin, by Merck Group (30 mentions)
Naringenin, by Merck Group (30 mentions)
Gallic acid, by Merck Group (29 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (28 mentions)
Luteolin, by Merck Group (26 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (26 mentions)
Kaempferol, by Merck Group (25 mentions)
Biochanin a, by Merck Group (24 mentions)
Rutin, by Merck Group (24 mentions)
Formononetin, by Merck Group (22 mentions)
Glycitein, by Merck Group (20 mentions)
Catechin, by Merck Group (19 mentions)
Caffeic acid, by Merck Group (19 mentions)
P coumaric acid, by Merck Group (18 mentions)
Epicatechin, by Merck Group (17 mentions)
409 protocols using genistein
1
Genistein Treatment of Uterine Cells
2
Genistein and Calcitriol Storage
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Genistein (4′,5,7-Trihydroxyisoflavone) and calcitriol (1α,25-Dihydroxycholecalciferol) were purchased from Sigma Aldrich (Germany) were stored at -20°C in the dark as single-used aliquots of concentrated stock solutions in dimethylsulfoxide (DMSO, for Genistein) or ethanol (EtOH, for calcitriol).
3
Genistein-Induced Apoptosis in Cell Lines
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Dulbecco's modified Eagle's Medium (DMEM), 100× penicillin and streptomycin, and fetal bovine serum (FBS) were acquired from Lonza (Verviers, Belgium). The Annexin-V–PI Apoptosis Kit and TUNEL Apo-Green Detection Kit were purchased from Biotool (Shanghai, China). DMSO and genistein were both purchased from Sigma-Aldrich (Shanghai, China), and a stock solution (200 mM) of genistein was prepared in DMSO. Matrigel™ was acquired from Invitrogen (Karlsruhe, Germany), and WST-8 and the Phosphatase and Protease Inhibitor Cocktail Set I were acquired from Biotool. First and second antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Detailed information regarding the antibodies we used are listed in Table 1 . The enhanced chemiluminescence (ECL) detection system was obtained from Biotool.
4
Establishing Genistein-Resistant Pancreatic Cancer Cells
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All cells were maintained in a 37°C incubator containing humidified air enriched with 5% CO2. Panc-1 and PaCa were continuously exposed to genistein (Sigma-Aldrich, St. Louis, MO, United States) to construct genistein-resistant cells. In brief, cells were exposed to different doses of genistein. Initially, the cells were exposed to 5 μM genistein for 48 h, once a week. When cells entered an exponential growth phase, the concentration of genistein was gradually increased till 100 μM. After 6 months of genistein treatment, the drug-resistant cells were sub-cultured and treated with 20 μM genistein every month to maintain the resistance. In the subsequent experiments, the genistein-resistant Panc-1 and PaCa cells were placed in a genistein-free condition for 14 days before use.
5
Myocarditis Cell Injury Model
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Rat cardiomyocytes H9c2 cells were acquired from American Type Culture Collection (ATCC), and incubated in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin in a humidified incubators at 37°C with 5% CO2. The cells injury model of myocarditis was formed by H9c2 cells with lipopolysaccharide (LPS, 10 μg/ml) stimulation for 12 h.
Sequences of si-con and si-Myc provided by Genepharma (Shanghai, China) were transfected into H9c2 cells with the help of Lipofectamine 2000 (Thermo Fisher Scientific, U.S.A.) to regulate the expression of Myc. Genistein was gained from Sigma and dissolved with dimethylsulfoxide (DMSO) to 10 µM. After pretreatment with 10 µM Genistein for 12h, H9c2 cells were stimulated with 10 μg/ml LPS for 12 h. H9c2 cells in the control group were cultured with equal concentration of DMSO.
Sequences of si-con and si-Myc provided by Genepharma (Shanghai, China) were transfected into H9c2 cells with the help of Lipofectamine 2000 (Thermo Fisher Scientific, U.S.A.) to regulate the expression of Myc. Genistein was gained from Sigma and dissolved with dimethylsulfoxide (DMSO) to 10 µM. After pretreatment with 10 µM Genistein for 12h, H9c2 cells were stimulated with 10 μg/ml LPS for 12 h. H9c2 cells in the control group were cultured with equal concentration of DMSO.
6
Genistein and AG1024 Effects on Prostate Cancer Cells
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The human PCa cell lines PC3 and DU145 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were maintained in RPMI-1640 medium (Gibco-BRL; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco-BRL; Thermo Fisher Scientific) at 37°C under 5% CO2 in a humidified incubator. For cell treatment, PC3 and DU145 cells were cultured in RPMI-1640 medium supplemented with genistein (purity >98%, Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and AG1024 (Sigma-Aldrich; Merck KGaA) at series concentrations. genistein (0, 10, 20, 30, 50 and 100 µM) and AG1024 (10 µM) (5 (link)) were dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich; Merck KGaA). X-ray irradiation was delivered using an X-6 MV photon linear accelerator (Varian Associates Inc., Palo Alto, CA, USA) at room temperature with a dose rate of 2 Gy/min (25 (link)). Cells treated with DMSO were considered as negative control for irradiation or drug treatment.
7
Genistein Attenuates Hepatic I/R Injury
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The present study was performed at Istanbul University Aziz Sancar Experimental Medicine Research Institute after the approval of Istanbul University Animal Experiments Local Ethics Committee (approval date and number: 10.08.2017/ 303098).
In the study, 32 male Wistar Albino rats weighing 200-250 g were used. Rats were housed in groups of four in cages with a temperature of 21 ± 1 °C and 12 hours of light / dark cycles, fed with standard laboratory diet and tap water ad libitum. The rats were randomized into 4 groups as follows:
Group I: Sham group: In this group, the rats underwent laparotomy without experimental hepatic ischemia-reperfusion injury and without medical treatment. Blood and liver tissue samples were taken.
Group II: Ischemia-Reperfusion (I/R) Group: No treatment was given to the rats in this group. After laparotomy, hepatic pedicle was occluded with an atraumatic vascular clamp for 15 minutes. Blood and liver tissue samples were taken after 20 minutes of reperfusion.
Group III: Genistein group: In this group, the rats were given 1 mg/kg of subcutaneous Genistein (Sigma Aldrich, USA) injection 24 hours and an hour before the procedure. Hepatic ischemia-reperfusion model was not applied to the rats. Laparotomy was performed under general anesthesia. Blood and liver tissue samples were obtained immediately after laparotomy.
In the study, 32 male Wistar Albino rats weighing 200-250 g were used. Rats were housed in groups of four in cages with a temperature of 21 ± 1 °C and 12 hours of light / dark cycles, fed with standard laboratory diet and tap water ad libitum. The rats were randomized into 4 groups as follows:
Group I: Sham group: In this group, the rats underwent laparotomy without experimental hepatic ischemia-reperfusion injury and without medical treatment. Blood and liver tissue samples were taken.
Group II: Ischemia-Reperfusion (I/R) Group: No treatment was given to the rats in this group. After laparotomy, hepatic pedicle was occluded with an atraumatic vascular clamp for 15 minutes. Blood and liver tissue samples were taken after 20 minutes of reperfusion.
Group III: Genistein group: In this group, the rats were given 1 mg/kg of subcutaneous Genistein (Sigma Aldrich, USA) injection 24 hours and an hour before the procedure. Hepatic ischemia-reperfusion model was not applied to the rats. Laparotomy was performed under general anesthesia. Blood and liver tissue samples were obtained immediately after laparotomy.
8
Whole-Cell Patch-Clamp Recordings of CFTR Activity
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Patch-clamp experiments were performed with an automatic electrophysiology workstation (Port-a-Patch, Nanion Technologies GmbH, Hambourg, Germany) coupled to an external amplifier unit HEKA EPC-10 [37 (link),38 (link)]. Whole-cell recordings were performed with treated or non-treated cells with the MBTP1 inhibitor. All measurements were obtained at room temperature. The voltage clamp protocol was carried out between −80 and +80 mV (10 mV per steps) with a holding membrane potential of −80 mV. The following buffers were used to suspend the cells: 140 mM NaCl, 2 mM CaCl2, 1 mM MgCl2, 10 mM Hepes (pH 7.4), 5 mM D-glucose monohydrate, 298 mOsm. The internal buffers were 50 mM CsCl, 10 mM NaCl, 60 mM Cs-Fluoride, 20 mM EGTA, 10 mM Hepes/CsOH, 5 mM Mg-ATP; pH 7.2; 285 mOsmol. CFTR’s activators (forskolin, 10 µM and genistein, 30 µM; Sigma Aldrich, Saint-Quentin Fallavier, France) and inhibitor (CFTRinh172, 10 µM; Sigma Aldrich) were added to activate or inhibit its activity, respectively.
9
Identification of Plant Defense Compounds
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Genistein, genistin, daidzin, daidzein, jasmonic acid (JA), coronatine, benzo(1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (acibenzolar S-methyl), and benzo(1,2,3) thiadiazole-7-carbothioic acid (acibenzolar acid) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Coumestrol, 2′-hydroxyGenistein, dalbergioidin, phaseollin, phaseollidin, kievitone, and phaseollinisoflavone were obtained and identified according to Durango et al. (2002) . Methyl jasmonate (MeJA), cis-jasmone and 2-carboxy cinnamic acid were from Alfa-Aesar Co. (Ward Hill, MA, USA). Methanol, ethyl acetate, NaOH, silica gel and all other chemicals were purchased from Merck KGaA (Darmstadt, Germany). 1-oxoindanoyl-amino acid conjugates were prepared from 2-carboxy cinnamic acid according to Krumm et al. (1995) (link) and Botero et al. (2021) .
10
Quantification of Isoflavones and Antioxidant Evaluation
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Reference standards of daidzein, genistein, puerarin, formononetin, and biochanin A with a purity of ≥98% were purchased from Sigma Aldrich (St. Louis, MO, USA) and used without further purification. Methanol (99.9%, for HPLC gradient), formic acid (98.0–100%, Puriss., for meeting analytical specifications of DAC and FCC), acetic acid (99.8%, for HPLC), and acetonitrile (99.9%, HPLC) were purchased from Sigma Aldrich. Choline chloride (99%; pharmaceutical grade) was purchased from Acros Organics, Geel, Belgium. Citric acid (99%, food-grade) was purchased from Sigma Aldrich. Quercetin, gallic acid, and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich. Ethyl acetate and ethanol were purchased from Himreaktivsnab company, Ufa, Russia. All other reagents and chemicals used in this study were of analytical grade. An ultrasonic cleaner and laboratory centrifuge Elma PE-6926 with a 10 × 5 mL rotor were used for the extraction process. A spectrophotometer Shimadzu-UV 1800 (Chiyoda-ku, Tokyo, Japan) was used as an analytical tool for the evaluation of total polyphenol content and antioxidant activity. A hot oven (Dry Oven UN55, Memmert, Schwabach, Germany) was used to dry the samples. In addition, HPLC-ESI-HRMS was used to quantify isoflavone content.
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