Black soybeans were purchased from NongHyup (Yeoryang, South Korea) and stored at 5 ℃ before the preparation of soybean milk. For the analysis of volatile flavor compounds, C7 − C40n-alkane standard and a 50/30 μm divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS) solid-phase microextraction (SPME) fiber were purchased from Supelco (Bellefonte, PA, USA). Butyrophenone was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) as an internal standard. For the analysis of isoflavones, daidzein, genestein, glycitein, daidzin, and genistin (Sigma-Aldrich Chemical Co.) were used as standards. HPLC-grade water, acetonitrile, acetic acid, and methanol were purchased from J.T. Baker (Philipsburg, NJ, USA). All chemicals and reagents used in the study were of analytical grade.
Genestein
Manufactured by Merck Group
Sourced in United States, United Kingdom
Genestein is a laboratory equipment product manufactured by the Merck Group. It is a device designed for the analysis and manipulation of genetic material. The core function of Genestein is to enable researchers to perform various genetic-related experiments and procedures in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Daidzein, by Merck Group (2 mentions)
Sodium butyrate, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Daidzin, by Merck Group (1 mentions)
Glycitein, by Merck Group (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Methanol, by Avantor (1 mentions)
Genistin, by Merck Group (1 mentions)
Acetic acid, by Avantor (1 mentions)
Hplc grade water, by Avantor (1 mentions)
C7 c40n alkane standard, by Merck Group (1 mentions)
Butyrophenone, by Merck Group (1 mentions)
Chlorpromazine, by Merck Group (1 mentions)
Carbenicillin disodium salt, by Merck Group (1 mentions)
Expifectamine 293 transfection kit, by Thermo Fisher Scientific (1 mentions)
Filipin, by Merck Group (1 mentions)
Valproic acid sodium salt vpa, by Merck Group (1 mentions)
Methyl β cyclodextrin, by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Tissue culture flasks, by BD (1 mentions)
4 protocols using genestein
1
Volatile Flavor and Isoflavone Analysis of Black Soybean Milk
2
Transient Transfection Reagents and Protocols
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DMSO (CAT#02650), sodium butyrate (CAT#156547), valproic acid sodium salt (VPA, CAT#P4543), carbenicillin disodium salt (CAT#C1389), Genestein (CAT# G6649), chlorpromazine (CAT# C8138), Filipin (CAT# F9765), and methyl-β-cyclodextrin (CAT# C4555) were purchased from Sigma-Aldrich (Burlington, MA, USA). sodium butyrate (CAT#M19-137) was from EMD Millipore (Billerica, MA, USA). Polyethyleneimine Max powder (CAT#24765-1, Polysciences Inc., Warrington, PA, USA) was dissolved in cell culture grade sterile water, adjusted to pH 7.0, sterile-filtered to the final stock solution of 1.5 mg/mL, aliquoted and stored at −20 °C. Tryptone N1 powder (CAT#19553, Organotechnie, La Courneuve, France) was dissolved into Expi293 expression medium to a final concentration of 20% (w/v) and sterile filtered. ExpiFectamine™ 293 Transfection Kit (CAT#A14524) and ExpiFectamineTM CHO Transfection Kit (CAT#A29131) were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
3
Genistein Cytotoxicity Evaluation in Cells
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In passage 3, cells were seated in 96 well plates and treated with variety concentrations of genestein (Sigma, UK) (0, 0.01, 0.004, 0.002 and 0.001M) and incubated for 24, 48 and 72 hours. After incubation times 5mg/ml MTT (Dimethylthiazolyldiphenyltetrazolium Bromide) was added to the cells and incubated for 4-5 hours at 37ºC in humidified CO2 incubator. Afterwards, MTT liquid was completely removed and kept up in the dark place and then DMSO (Merck, Germany) was added to each well, incubated for 24 hours and the absorbance of the plate was read at 495 nm.
4
Caco-2 Cell Response to Isoflavones
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The Caco-2 cell line (ICLC HTL 97023-Interlab Cell Line Collection, Genoa, Italy) was grown in MEM medium supplemented with 10% fetal bovine serum (FBS, UE approved origin), 100 U/mL penicillin, 100 μg/mL of streptomycin, L-glutamine (2 mM), 1% nonessential amino acids (NEEA), referred to as complete medium (all reagents were purchased from Life Technologies-Invitrogen, Milan, Italy).
Cultures were maintained at 37° C in a humidified atmosphere containing 5% CO2 and expanded in tissue culture flasks (75 cm 2 , BD Bioscences, USA), changing the medium daily. The cells were seeded in 6-well cell culture plates at 5 x 10 5 cells/well and cultured to reach 80% confluency.
For the experiments, cells were treated with Salmonella enterica serotype typhimurium LPS (Sigma). Preliminary experiments were performed in order to establish the optimal dose (1 g/mL) of LPS and time of exposure to LPS (48 h). Before LPS stimulation, some wells were pre-treated with different concentrations (5, 10, 50 μM) of daidzein (4′,7-dihydroxyisoflavone, Sigma), genestein (4′, 5, 7-trihydroxyisoflavone, Sigma) and equol (dihydroxyisoflavone, Sigma) or with different amounts (10, 30 and 50 l) of the commercial (bovine or soy) milks. After 1h of incubation at 37°C, cell cultures were then stimulated with endotoxin as previously indicated. Untreated cells were used as control.
Cultures were maintained at 37° C in a humidified atmosphere containing 5% CO2 and expanded in tissue culture flasks (75 cm 2 , BD Bioscences, USA), changing the medium daily. The cells were seeded in 6-well cell culture plates at 5 x 10 5 cells/well and cultured to reach 80% confluency.
For the experiments, cells were treated with Salmonella enterica serotype typhimurium LPS (Sigma). Preliminary experiments were performed in order to establish the optimal dose (1 g/mL) of LPS and time of exposure to LPS (48 h). Before LPS stimulation, some wells were pre-treated with different concentrations (5, 10, 50 μM) of daidzein (4′,7-dihydroxyisoflavone, Sigma), genestein (4′, 5, 7-trihydroxyisoflavone, Sigma) and equol (dihydroxyisoflavone, Sigma) or with different amounts (10, 30 and 50 l) of the commercial (bovine or soy) milks. After 1h of incubation at 37°C, cell cultures were then stimulated with endotoxin as previously indicated. Untreated cells were used as control.
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