To prepare the injection solution, 6-shogaol (Merck) was weighed and dissolved in the base solution of saline with 0.2% DMSO to obtain a final concentration of 0.12 mg/mL. All the injection solutions (70% methanol extract, 80% ethanol extract, 100% DMSO extract, and 6-shogaol) were kept in dark containers at room temperature.
6 shogaol
Manufactured by Merck Group
Sourced in United States, Germany
6-shogaol is a chemical compound found in ginger root. It is a common research tool used in laboratory settings for various scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
6 gingerol, by Merck Group (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Curcumin, by Merck Group (3 mentions)
8 gingerol, by Merck Group (3 mentions)
6 paradol, by Merck Group (2 mentions)
Lc ms grade acetonitrile, by Merck Group (2 mentions)
Milli q water system, by Merck Group (2 mentions)
10 gingerol, by Merck Group (2 mentions)
Plumbagin, by Merck Group (2 mentions)
Fitc annexin 5 apoptosis detection kit, by BD (2 mentions)
Piperine, by Merck Group (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Methanol, by Merck Group (2 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Opti mem, by Capricorn (1 mentions)
Dulbecco s modified eagle s medium, by Capricorn (1 mentions)
Protocatechuic acid, by Merck Group (1 mentions)
Penicillin streptomycin, by Capricorn (1 mentions)
Fetal bovine serum (fbs), by Capricorn (1 mentions)
Dibenzylideneacetone, by Merck Group (1 mentions)
23 protocols using 6 shogaol
1
Preparation of 6-shogaol Injection Solution
2
Alzheimer's Biomarker Evaluation and Antioxidant Analysis
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Dimethyl sulfoxide (DMSO) was purchased from BioLife Solutions (Bothell, WA, USA). Protease inhibitor (PI) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Opti-MEM, Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), and penicillin/streptomycin were obtained from Capricorn Scientific (Ebsdorfergrund, Germany). An enzyme-linked immunosorbent assay (ELISA) kit for Aβ40/42 was obtained from IBL (Kunma, Japan). The ELISA kit for tumor necrosis factor alpha (TNF-α), interleukin (IL-1β), and interleukin (IL-6) was purchased from R&D Systems (Minneapolis, MN, USA). The AChE assay kit and the Randox Total Antioxidant Status (TAS) kit were purchased from BioVision (Milpitas, CA, USA) and Randox (Crumlin, UK). Cell viability assay reagent (CCK-8) was purchased from Dogenbio Co. Ltd. (Seoul, Korea). Mouse anti-Aβ17-26 monoclonal antibody (4G8) was purchased from BioLegend (San Diego, CA, USA). Aβ1-42 was purchased from American Peptide Company, Inc. (Sunnyvale, CA, USA). Curcumin, 6-shogaol and protocatechuic acid were purchased from Sigma (St. Louis, MO, USA). All other reagents were of highest purity grade.
3
Curcumin, Dibenzylideneacetone, and 6-Shogaol Assay
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Curcumin (cat #C7727), dibenzylideneacetone (cat#246425) and 6-shogaol (cat #39303) were purchased from Sigma-Aldrich.
4
Evaluating Pharmacological Inhibitors of ROS
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Compounds were obtained as follows: diphenyleneiodonium (Nox or ROS inhibitor, DPI, Sigma Aldrich, St Louis, MO, USA), N-acetylcysteine (a ROS inhibitor, NAC, Sigma Aldrich, St Louis, MO, USA), Z-VAD-FMK (caspase inhibitor, Sigma Aldrich, St Louis, MO, USA), 6-shogaol (Sigma Aldrich, St Louis, MO, USA), and thapsigargin (ER stress inducer, TG, Millipore, Bedford, MA, USA).
5
Anti-inflammatory Effects of 6-Shogaol
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6-Shogaol (Sigma-Aldrich, St.Louis, MO, USA), Fetal bovine serum (FBS) (Thermoscientific, USA), RPMI1640 medium (Gibco, Grand Island, NY, USA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), polyvinylidenedifluoride (PVDF) membranes and enhanced chemiluminescence (ECL) kit (Invitrogen) were used in the study. ELISA kits for determining cytokines-TNF-α, IL-1β and IL-6 were purchased from Biolegend (San Diego, CA, USA). Buffers used in Western blotting analysis were procured from Beyotime Institute of Biotechnology (Beijing, China). Antibodies against VEGF, VEGF-A, VEGFR-2 and COX-2 were procured from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxidase-labelled IgG secondary antibodies, PGE2, TNF-α, NF-κB p65, IκBα, p-IκBα, p-IKKβ, IKKβ, p-IKKα, IKKα and β-actin were purchased from Santa Cruz Biotechnology (Texas, USA).
6
DPPH-Based Antioxidant Activity Evaluation
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Antioxidant activity of the extracts were estimated using DPPH in vitro method Mensor et al. [18] (link). Test samples were dissolved separately in methanol to get test solution of 1 mg/mL. Series of extract/ pure components (6-gingerol and 6-shogaol; purchased from (Sigma-Aldrich GmbH, Germany) solutions of different concentrations (100, 200, 400, 600, 800 and 1000 µg/mL) were prepared by diluting with methanol. Assays were performed in 96-well, microtiter plates. 140 µL of 0.6 × 10−6 mol/L DPPH were added to each well containing 70 µL of sample. The mixture was shaken gently and left to stand for 30 min in dark at room temperature. The absorbance was measured spectrophotometrically at 517 nm using Cecil-Elect Spectrophotometer. Blank was done in the same way using methanol and sample without DPPH and control was done in the same way but using DPPH and methanol without sample. Ascorbic acid was used as reference antioxidant compound. Every analyse is done in triplicate. The ability to scavenge DPPH radical was calculated by the following equation: where
Abssample is the absorbance of DPPH radical + sample;
Absblank is the absorbance of sample + methanol;
Abscontrol is the absorbance of DPPH radical + methanol.
7
Fluorescent Lipophilic Dyes and Immune Cell Markers
8
Fibrosarcoma and Periodontal Fibroblast Culture
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The fibrosarcoma cell line (HT1080) was obtained from American Type Culture Collection (ATCC-CCL-12), and the human periodontal-derived ligament fibroblast (HPdLF) line from Lonza Ref. CC-7049 (Walkersville, MD, USA). Both cells lines were maintained in complete DMEM Dulbecco’s modified Eagle medium that was supplemented with 10% fetal bovine serum (Lonza, Walkersville, MD, USA) and a 1% antibiotic–antimycotic cocktail, and at 37 °C and 5% CO2. 6-Shogaol was obtained from Sigma-Aldrich (St. Louis, MO, USA) [13 (link)].
9
UHPLC Analysis of Ginger Bioactives
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The Agilent Model 1200 UHPLC system was used. The 6-gingerol and 6-shogaol were separated on an Agilent C18 (4.6 × 250 mm, 5 μm) reversed-phase column by gradient elution using: (A) water and (B) acetonitrile. The detector wavelength was set at 280 nm. The flow rate and injection volume were 1 mL/min and 20 µL, respectively. Identification of the compounds was achieved by comparison of retention times with standards (6-gingerol and 6-shogaol), UV spectra and UV absorbance ratios after co-injection of samples and standards. 6-gingerol (≥98%, CAS Registry number: 23513-14-6), 6-shogaol (≥90%, CAS Registry number: 555-66-8), 8-gingerol (≥95%, CAS Registry number: 23513-08-8), were purchased from Sigma-Aldrich, Selangor, Malaysia and 8-shogaol (≥98%, CAS Registry number: 36700-45-5) was purchased from Chem Faces, Wuhan, China. System suitability requirements: at least five replicate injections of 6-and 8-gingerol and shogaol were performed; the requirements of the system suitability parameters are: (1) symmetry factor is not more than 1.5 (2) percentage of relative standard deviation of the retention time for 6-gingerol and 6-shogaol standards is not more than 2.0%.
10
HPLC Analysis of Ginger Bioactive Compounds
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The 6-gingerol and 6-shogaol were analyzed by high-performance liquid chromatography (HPLC) using a JASCO HPLC system (Tokyo, Japan), according to a method by Moon et al. [10 (link)]. Standard 6-gingerol and 6-shogaol were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Total saponin content of each extract was determined using the method described by Moon et al. [10 (link)]. The ginsenoside Re (Wako Chem. Co., Osaka, Japan) was used as a reference standard.
Total saponin content of each extract was determined using the method described by Moon et al. [10 (link)]. The ginsenoside Re (Wako Chem. Co., Osaka, Japan) was used as a reference standard.
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