Minimum essential medium-α (α-MEM) and penicillin/streptomycin (P/S) were obtained from Gibco (Gaithersburg, MD, USA). The CCK-8 assay was obtained from Dojindo Molecular Technologies, Inc. (Kumamoto, Japan). Fetal bovine serum (FBS) was supplied by Atlas Biologicals (Fort Collins, CO, USA). Bicinchoninic acid (BCA) protein assay kit, β-glycerophosphate, ascorbic acid, dimethyl sulfoxide (DMSO) and amygdalin were obtained from Sigma Aldrich (St. Louis, MO, USA). Dulbecco’s phosphate-buffered saline was supplied by Welgene, Inc. (Daejeon, Republic of Korea). Anti-RUNX2, anti-BMP-2 and anti-SMAD1/5/9 antibodies were supplied by Abcam (Cambridge, UK). Anti-t-p38 and anti-p-p38 antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Secondary antibodies were supplied by Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA). PCR primers were supplied by Genotech (Daejeon, Republic of Korea). The SuperScript II Reverse transcription kit and SYBR-Green solution were purchased from Invitrogen (Carlsbad, CA, USA). Taq polymerase was purchased from Kapa Biosystems (Woburn, MA, USA). The OCN (cat. no: LS-F12230) and the OSN (cat. no: LS-F27540) ELISA kit was obtained from LSbio (Seattle, WA, USA).
Dulbecco s phosphate buffered saline
Manufactured by Welgene
Sourced in United States
Dulbecco's phosphate-buffered saline (DPBS) is a balanced salt solution commonly used in cell culture and biomedical research. It is a sterile, isotonic buffer that maintains the pH and osmotic balance of cell media. DPBS is formulated to support the physiological environment of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Welgene (3 mentions)
Penicillin streptomycin antibiotic solution, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Atlas Biologicals (1 mentions)
Superscript 2 reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
α minimum essential medium α mem, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid bca protein assay kit, by Merck Group (1 mentions)
Taq polymerase, by Roche (1 mentions)
Cck 8 assay, by Dojindo Laboratories (1 mentions)
Sybr green solution, by Thermo Fisher Scientific (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Amygdalin, by Merck Group (1 mentions)
Freezone plus, by Labconco (1 mentions)
B1000, by Thermo Fisher Scientific (1 mentions)
Las 3000 imaging system, by Fujifilm (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Laemli sample buffer, by Bio-Rad (1 mentions)
17 protocols using dulbecco s phosphate buffered saline
1
Osteogenic Differentiation Protocol for Stem Cells
2
Quantitative Analysis of hIL-10 Expression
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For hIL-10 detection, the supernatant of PK(15) and PK(15)-hIL10 was collected and centrifuged at 300 g for 10 minutes. The samples were lyophilized using Freezone Plus (Labconco, 7960040) and the protein concentration was determined with the Bradford assay (Bio-rad, 500-0006). Proteins (5 μg/well) were added 2 X Laemli sample buffer (Bio-rad, 1610737) and reduced. Samples were separated by electrophoresis (Life technologies™, B1000) on 4 %–12% Bis-Tris polyacrylamide gels (Invitrogen, NW04120BOX), and the bands were transferred to nitrocellulose membrane (Bio-rad, 1620115). The membranes were blocked in Dulbecco’s Phosphate Buffered Saline (Welgene, LB001-02) with 0.1% tween 20 (Sigma, P9416)/5% skim milk (BD, 232100) for 1 hour at room temperature. Primary and secondary antibodies were diluted at 1:1000 and 1:5000, respectively for blotting. Quantitation and imaging of western blots were done using LAS 3000 imaging system (Fuji), following the manufacturer's instructions.
3
Culturing and Identification of SARS-CoV-2 using Vero E6 Cells
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Vero E6 cells were used for the culturing and identification of SARS-CoV-2. All cell culture work of SARS-CoV-2 virus was performed in a laboratory with Biosafety Level 3 (BL3) facility (Health and Environment Research Institute of Gwangju City). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and a 1× penicillin–streptomycin antibiotic solution (Gibco, Thermo Fisher Scientific Inc., Korea) in an atmosphere containing 5% CO2 at 37°C. The swab samples were obtained by means of the UTM kit with 1 mL of the viral transport medium (Noble Bio, Korea), or the samples collected into collection tubes were diluted with 1 mL of Dulbecco’s phosphate-buffered saline (Welgene, Korea) and thereafter inoculated into a monolayer of the cultured Vero cells. The inoculated culture was examined daily for cytopathic effects, similar to the procedures used for SARS-CoV and MERS-CoV in other studies (32 (link), 33 (link)).
4
Antibody Labeling and Flow Cytometry
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The cells were detached and resuspended in Dulbecco's phosphate-buffered saline (DPBS; Welgene, Dae-gu, Korea) and 2% FBS. About 1.0 × 105 cells were applied with antibodies for 30 minutes on ice. The antibodies are listed in Supplementary Table 1 available online at https://doi.org/10.1155/2017/8085462 . After washing, the fluorescence intensity was determined by a FACS Calibur (Becton Dickinson, San Jose, CA, USA). For the analysis of data, we used FLOWJO software (Tree Star Inc., Ashland, OR, USA).
5
Cytotoxicity Assay with 6-Gingerol
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Dulbecco’s phosphate-buffered saline (DPBS) and Roswell Park Memorial Institute-1640 (RPMI-1640) medium were obtained from WELGENE (Gyeongsan, Korea). 6-Gingerol, DMSO, and MTT were purchased from Sigma (St. Louis, MO, USA).
6
Cell Culture Reagents and Treatments
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Carvedilol, 3,4-dihydroxy-L-phenylalanine (L-DOPA), cholera toxin (CT), and 12-O-tetradecanoylphorbol-13-acetate (TPA) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM) and Dulbecco’s phosphate-buffered saline were purchased from WelGENE (Daegu, Korea). Fetal bovine serum (FBS), antibiotic-antimycotic, and trypsin-EDTA were purchased from Gibco (Grand Island, NY, USA). Medium 254 (Cascade Biologics, Portland, OR, USA) and FSK ([3R-(3α,4aβ,5β,6β,6aα,10α,10aβ,10bα)]-5-(Acetyloxy)-3-ethenyldodecahydro-6,10,10b-trihydroxy-3,4a,7,7,10a-pentamethyl-1H-naphtho[2,1-b]pyran-1-one) were purchased from Tocris Bioscience (Bristol, UK).
7
Molecular Mechanisms of Insulin Signaling
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Telmisartan and losartan were purchased from Cayman Chemical (Ann Arbor, MI, USA). Fimasartan was a kind gift from Boryung Pharmaceutical (Seoul, Korea). D-Glucose, GW9662, sodium lactate, HEPES, dimethyl sulfoxide (DMSO), and bovine insulin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against Akt, p-Akt-Ser473, GSK3β, p-IRS-1-Ser302, p-IRS-Ser307, p-IRS-Ser318, p-IRS-Ser1101, p-IRS-Tyr632, p-IRS-Tyr896, IRS-1, and actin were purchased from Cell Signaling Technology (Boston, MA, USA). Antibodies against glucose-6-phosphatase α (G6Pase-α), phosphoenolpyruvate carboxykinase (PEPCK), and PKCζ, p-PKCζ-Thr410 were purchased from Santa Cruz Biotechnology (La Jolla, CA, USA). Antibodies against p-GSK3β-Ser9 and hemagglutinin (HA) were obtained from BD Biosciences (San Jose, CA, USA) and Covance Inc. (Emeryville, CA, USA), respectively. Low or high glucose Dulbecco’s modified Eagle’s medium (DMEM) and Dulbecco’s phosphate-buffered saline were purchased from Welgene Inc. (Gyeongsan, Korea). Sodium pyruvate, fetal bovine serum (FBS), penicillin and streptomycin antibiotics, L-glutamine, and trypsin-EDTA solution were purchased from Gibco-BRL (Gaithersburg, MD, USA). Plasticware for cell culture was purchased from Corning Inc. (Oneonta, NY, USA) or SPL Life Sciences (Pocheon, Korea). All other chemicals were of the purest analytical grade.
8
Generation of Immune Cell Cultures
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Phorbol-12-myristate 13-acetate (PMA) was purchased from Cayman Chemical (Ann Arbor, MI, USA). Dulbecco’s phosphate-buffered saline (DPBS), RPMI-1640 culture medium, and fetal bovine serum (FBS) were purchased from Welgene, Inc. (Daegu, Republic of Korea). Dopamine hydrochloride, RGD, and dexamethasone (Dex) were purchased from Sigma-Aldrich (St. Louis, MO, USA). SU-8 3005 and SU-8 developer were purchased from MicroChem (Round Rock, TX, USA). PDMS prepolymer (Sylgard 184) and a curing agent were purchased from Dow Corning (Midland, MI, USA). Norland Optical Adhesive (NOA) 63 was purchased from Norland Products (Jamesburg, NJ, USA). Tris-HCl was purchased from Biosesang (Seongnam, Republic of Korea). Ethylenediaminetetraacetic acid (EDTA) was purchased from Amresco Inc. (Solon, OH, USA). Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon (IFN)-γ were purchased from R&D Systems (Minneapolis, MN, USA). Pluronic F-127 solution and antibiotic–antimycotic solution were purchased from Invitrogen Corp. (Waltham, MA, USA). 1,25-dihydroxy vitamin D3 (VitD3) was purchased from Toronto Research Chemicals Inc. (Toronto, ON, Canada).
9
Characterization of Bacterial Pellicle Composition
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The harvested 3- to 4-day-old pellicles were washed with Dulbecco’s phosphate-buffered saline (Welgene) followed by treatment with 0.86 units (U)/mL 1,4-(1,3:1,4)-β-D-glucan 4-glucanohydrolase (v/v) (Calbiochem-Novabiochem), 0.1% proteinase K (v/v) (Sigma-Aldrich), or 0.1% RNase-free DNase I (v/v) (Qiagen) as described previously [26 (link), 27 (link)]. After overnight incubation at 37°C, the turbidity of the degraded pellicles was measured as the optical density at 600 nm (OD600) using an Eppendorf BioSpectrometer kinetic.
10
Culturing and Identification of SARS-CoV-2 using Vero E6 Cells
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Vero E6 cells were used for the culturing and identification of SARS-CoV-2. All cell culture work of SARS-CoV-2 virus was performed in a laboratory with Biosafety Level 3 (BL3) facility (Health and Environment Research Institute of Gwangju City). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and a 1× penicillin–streptomycin antibiotic solution (Gibco, Thermo Fisher Scientific Inc., Korea) in an atmosphere containing 5% CO2 at 37°C. The swab samples were obtained by means of the UTM kit with 1 mL of the viral transport medium (Noble Bio, Korea), or the samples collected into collection tubes were diluted with 1 mL of Dulbecco’s phosphate-buffered saline (Welgene, Korea) and thereafter inoculated into a monolayer of the cultured Vero cells. The inoculated culture was examined daily for cytopathic effects, similar to the procedures used for SARS-CoV and MERS-CoV in other studies (32 (link), 33 (link)).
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