Approximately 1.0 × 105 Huh7 or PLC/PRF/5 cells per well were plated into a 6-well plate and, 24 h later, were treated with or without 5 μM zinc chloride (Wako) in serum-free medium. The cellular RNA was extracted by an RNeasy Mini Kit (Qiagen). The cDNA was synthesized with an RT2 First Strand cDNA Kit (Qiagen), according to the manufacturer’s protocol [10 (link)]. A TLR signaling pathway PCR array was purchased from Qiagen. A real-time PCR array based on the SYBR Green method was performed on a Quant Studio 3 real-time PCR system (Applied Biosystems, Foster, CA, USA). The cycling program was as follows: 95 °C for 10 min for 1 cycle, then 40 cycles of 95 °C for 15 s and 60 °C for 1 min. The housekeeping genes beta-2-microglobulin (B2M), hypoxanthine phosphoribosyltransferase 1 (HPRT1), ribosomal protein L13a (RPL13A), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and actin beta (ACTB) served as the internal controls. The data were analyzed by the Excel-based RT2 Profiler PCR Array Data Analysis software and website, which was provided by Qiagen (https://dataanalysis2.qiagen.com/pcr (accessed on 4 Juanury 2021)).
Zinc chloride
Manufactured by Fujifilm
Sourced in Japan
Zinc chloride is a chemical compound used in various laboratory applications. It is a colorless, crystalline solid with the chemical formula ZnCl2. Zinc chloride is a Lewis acid and is commonly used as a desiccant, flux, and reagent in chemical reactions.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Fujifilm (2 mentions)
Sodium hydroxide (naoh), by Fujifilm (2 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Rt2 first strand cdna kit, by Qiagen (1 mentions)
Manganese 2 chloride tetrahydrate, by Fujifilm (1 mentions)
Cobalt 2 chloride hexahydrate, by Fujifilm (1 mentions)
Copper 2 chloride dihydrate, by Fujifilm (1 mentions)
Nickel ii chloride hexahydrate, by Fujifilm (1 mentions)
Cycloheximide (chx), by Abcam (1 mentions)
Hydrogen peroxide, by Fujifilm (1 mentions)
Mercury 2 chloride, by Fujifilm (1 mentions)
Hydrochloric acid (hcl), by Fujifilm (1 mentions)
Acetic acid, by Fujifilm (1 mentions)
Chitosan, by Tokyo Chemical Industry (1 mentions)
Cadmium chloride 2.5 hydrate, by Fujifilm (1 mentions)
Sodium chloride (nacl), by Fujifilm (1 mentions)
Decane, by Fujifilm (1 mentions)
Ofloxacin, by Fujifilm (1 mentions)
1 octanol, by Fujifilm (1 mentions)
8 protocols using zinc chloride
1
Zinc Chloride Modulates TLR Signaling in Liver Cells
2
Synthesis of Metal-Chelated Antioxidants
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Lauric
acid (CH3(CH2)10COOH), magnesium(II)
chloride hexahydrate, manganese(II)
chloride tetrahydrate, cobalt(II) chloride hexahydrate, nickel(II)
chloride hexahydrate, copper(II) chloride dihydrate, zinc chloride,
and phosphate buffer (pH 6) were purchased from Wako Pure Chemical
Industries Ltd. (Osaka, Japan). Calcium(II) chloride dihydrate was
obtained from Sigma-Aldrich Co. LLC (St. Louis). Sodium hydroxide
was purchased from Kanto Chemical Co. Inc. (Kanagawa, Japan). DMPO
was purchased from Labotech Inc. (Tokyo, Japan).
acid (CH3(CH2)10COOH), magnesium(II)
chloride hexahydrate, manganese(II)
chloride tetrahydrate, cobalt(II) chloride hexahydrate, nickel(II)
chloride hexahydrate, copper(II) chloride dihydrate, zinc chloride,
and phosphate buffer (pH 6) were purchased from Wako Pure Chemical
Industries Ltd. (Osaka, Japan). Calcium(II) chloride dihydrate was
obtained from Sigma-Aldrich Co. LLC (St. Louis). Sodium hydroxide
was purchased from Kanto Chemical Co. Inc. (Kanagawa, Japan). DMPO
was purchased from Labotech Inc. (Tokyo, Japan).
3
Preparation of Chemical Compounds
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Hydrogen peroxide, mercury II chloride, and zinc chloride were purchased from Wako, Japan. Cycloheximide was purchased from Biovision, USA. These chemicals were dissolved in dimethyl sulfoxide (Wako) and diluted in culture medium at 0.1% vol/vol final concentration.
4
Synthesis of Imogolite Clay Nanotubes
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Chitosan and PLP were purchased from Tokyo Chemical Industry Co., Ltd. Acetic acid, sodium hydroxide, hydrochloric acid, zinc chloride, and cadmium chloride 2.5-hydrate were purchased from FUJIFILM Wako Pure Chemical Corp. Since the physical and chemical properties of synthetic imogolite using Farmer et al. reported method have been well investigated, we synthesized imogolite by his method.7 (link)
5
Characterization of Fluoroquinolone Derivatives
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Norfloxacin (NFLX), ofloxacin (OFLX), tosufloxacin tosylate monohydrate (TFLX), calcium dichloride, magnesium dichloride, iron(II) dichloride tetrahydrate, iron(III) chloride hexahydrate, aluminum(III) chloride hexahydrate, zinc chloride, lanthanum chloride heptahydrate, 1-octanol, sodium dihydrogen phosphate dihydrate, sodium chloride, sodium hydroxide, decane, 8 M NaOH, and 0.1 vol% trifluoroacetic acid-acetonitrile were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). Ciprofloxacin hydrochloride monohydrate (CPFX), levofloxacin (LVFX), enoxacin sesquihydrate (ENX), fleroxacin (FLRX), and prulifloxacin (PUFX) were purchased from TCI (Tokyo, Japan). Gatifloxacin (GFLX) and sparfloxacin (SPFX) were purchased from Funakoshi Co., Ltd (Tokyo, Japan). Moxifloxacin (MFLX) was purchased from Combi-Blocks Inc. (San Diego, USA). 2-Morpholinoethanesulfonic acid (MES) was purchased from Dojindo laboratories (Tokyo, Japan). 0.1 vol% trifluoroacetic acid-distilled water was purchased from Kanto chemical co., inc (Tokyo, Japan). Soybean lecithin was provided by Tsuji Oil Mills co., Ltd (Mie, Japan) (SLP-White, phosphatidylcholine (24-32 %), phosphatidylethanolamine (20-28 %), phosphatidylinositol (12-20 %), phosphatidic acid (8-15 %), and lysophosphatidylcholines (1-5 %) (based on the product information provided by the manufacturer)).
6
Huh7 Cell Transfection and HAV Infection Study
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Approximately 1.0 × 105 Huh7 cells were seeded on 6-well tissue culture plates (Iwaki Glass, Tokyo, Japan) 24 h prior to transfection. A total of 50 μM si-MAP2K3 or 50 μM si-C were transfected into Huh7 cells using Effectene transfection reagent (Qiagen), according to the manufacturer’s instructions [13 (link)]. After 24 h of transfection, the cells were infected with 0.01 MOI of HAV genotype IIIA HA11-1299 after the cells were washed with phosphate-buffered saline (PBS) twice. After 24 h of infection, the cells were washed with PBS and maintained in 5% FCS DMEM, with or without zinc chloride (Wako). After 48 h of infection, the cellular RNA was extracted in order to determine the HAV RNA [10 (link)].
7
Monitoring S. mutans pH Reduction
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A reaction mixture (0.5 mL) of S. mutans cells (30 mg) suspended in 2 mM PPB was prepared. Then, potassium fluoride (Wako Pure Chemical Industries, Osaka, Japan) or zinc chloride (Wako Pure Chemical Industries) was added to the S. mutans cells suspension. The final concentration of fluoride and zinc ions in the reaction mixture was set according to the concentrations detected from the interface between bacteria and WSG-TRR. After pre-incubation, 1% glucose was added to the mixture, and the reduction in pH was monitored using a pH electrode (model 9618S, HORIBA, Kyoto, Japan) and pH meter (F-72, HORIBA) at 37°C for 90 min.
8
Optimizing Cell Culture Conditions with DMEM, RNAlater, and TRIzol
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Dulbecco s modified Eagle s medium (DMEM) , RNAlater, and TRIzol were obtained from Life Technologies (Carlsbad, CA, USA) , and ReverTra Ace was purchased from Toyobo (Osaka, Japan) . AmpliTaq Gold 360 Master Mix was obtained from Applied Biosystems (Foster City, CA, USA) . The Human Free BDNF ELISA Kit was purchased from R&D Systems (Minneapolis, MN, USA) . The primers used in this study (Table 1) were obtained from GeneNet (Fukuoka, Japan) or Greiner Japan (Kanagawa, Japan) . Whole-wheat flour and wheat endosperm were purchased from Hara Farm (Kumamoto, Japan) , and wheat bran was obtained from YouTech (Hokkaido, Japan) . Magnesium chloride hexahydrate, zinc chloride, methanol, 3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide (MTT) , dimethyl sulfoxide (DMSO) , and other reagents were obtained from Wako Pure Chemical Industry (Osaka, Japan) .
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