Hexane, dichloromethane, ethyl acetate and dimethyl sulfoxide (DMSO) were purchased from FS Chemicals (Francfort, Germany) (analytical grade). RPMI 1640 was purchased from Nacalai Tesque (Kyoto, Japan). Fetal bovine serum, trypsin, streptomycin and penicillin were obtained from PAA Laboratories GmBH (Pasching, Austria). 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT), propidium iodide and RNAse A were purchased from Sigma (St. Loius, USA). Tissue culture flasks, 6-well plates and 96-well plates were obtained from TPP (Trasadingan, Switzerland). Annexin-V FITC Kit was obtained from eBioscience Inc. (San Diego, USA). Real Genomics Total RNA extraction kit (RBC Biosciences, Taiwan) and GenomeLab GeXP Start Kit (Beckman Coulter, USA) were also procured.
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt propidium iodide
Manufactured by Merck Group
Sourced in United States
3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) is a chemical compound used as a reagent in cell viability assays. Propidium iodide is a fluorescent dye that binds to DNA and is commonly used in cell biology applications to detect cell death.
Automatically generated - may contain errors
Lab products found in correlation
Rnase a, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by GeneTex (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Genomelab gexp start kit, by Beckman Coulter (1 mentions)
Annexin 5 fitc kit, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Nacalai Tesque (1 mentions)
Cell culture reagents, by Welgene (1 mentions)
Tris buffer, by Thermo Fisher Scientific (1 mentions)
Normal human osteoblasts, by Lonza (1 mentions)
Recombinant human opg, by Thermo Fisher Scientific (1 mentions)
Acridin orange, by Merck Group (1 mentions)
Agarose, by Bio-Rad (1 mentions)
Ubiquitin, by Abcam (1 mentions)
Aldh1a1, by Cell Signaling Technology (1 mentions)
P pkc, by Cell Signaling Technology (1 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Anti mouse, by Cell Signaling Technology (1 mentions)
14 protocols using 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt propidium iodide
1
Compound Screening and Cell-based Analysis
2
Reagents and Antibodies for Cell Experiments
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Cell culture reagents were purchased from Welgene (Daegu, Republic of Korea). Mouse monoclonal antibodies against cleaved poly (ADP-ribose) polymerase (PARP) were purchased from BD Biosciences (San Jose, CA, USA). Rabbit polyclonal antibodies against pAMPK (T172), AMPK, pmTOR (S2448), mTOR, p70S6K (T389), p70S6K, pS6, S6, and cleaved caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Primary antibodies against actin and SDHA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit polyclonal SDHB antibody and horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from GeneTex (Irvine, CA, USA). Primary antibodies against SDHB and SDHC were purchased from Abcam (Cambridge, UK). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), propidium iodide (PI), and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise specified.
3
Chitosan and Recombinant Human OPG on Periodontal Cells
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Low molecular weight (LMW), medium molecular weight (MMW) and high molecular weight (HMW) chitosan and human OPG protein (Recombinant Human OPG; PeproTech, Rocky Hill, New Jersey, USA) were used in this study. Tris buffer (5 mmol L−1, pH 7.5) and dimethyl sulfoxide (DMSO) (Fisher Scientific, Leics, UK) were used throughout the experiment. Normal, human periodontal ligament (NHPL) fibroblasts and normal, human osteoblasts were obtained from Lonza (Lonza Inc., Walkersville, MD, USA). Penicillin-streptomycin (Bioscience Ltd, Buckingham, UK), 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) propidium iodide and acridin orange (Sigma-Aldrich, St. Louis, MO, USA) were also used.
4
Stem Cell Proliferation and Phenotype
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Roswell Park Memorial Institute (RPMI) 1640 medium, penicillin/streptomycin, fetal bovine serum (FBS), phosphate-buffered saline (PBS), L-glutamine, and trypsin-EDTA were acquired from Gibco (Grand Island, NY, USA). Dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), propidium iodide (PI), Hoechst 33342, Triton X-100, bovine serum albumin (BSA), MG132, and paraformaldehyde were obtained from Sigma-Aldrich, Co. (St. Louis, MO, USA). Agarose was obtained from Bio-Rad Laboratories (Hercules, CA, USA). RIPA buffer and wortmannin were acquired from Cell Signaling Technology, Inc. (Danvers, MA, USA). The primary antibodies used in the experiments including β-Actin (#4970), Akt (#9272), phosphorylated Akt or p-Akt (#4060), c-Myc (#5605), PKC (#2056), p-PKC (#9375), GSK3β (#9323), p-GSK3β (#9832), mTOR (#2983), p-mTOR (#5536), Nanog (#4903), Oct4 (#2840), Sox2 (#3579), and ALDH1A1 (#36671) were acquired from Cell Signaling Technology (Danvers, MA, USA). The primary antibody ubiquitin (ab7780) was purchased from Abcam (Cambridge, UK). The respective secondary antibodies, anti-rabbit IgG (#7074), and anti-mouse (#7076) were also obtained from Cell Signaling Technology (Danvers, MA, USA).
5
Curcumin-Loaded PLGA Nanoparticles
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Poly (D, L-lactide-co-glycolide) (50:50) (PLGA-A, Purasorb Polymer PDLG 5002 A, Mw ~17,000) was kindly provided by Corbion Purac (The Netherlands). Eudragit S100 (Mw ~ 125,000 g/mol) was obtained from Evonik (India). Curcumin and polyvinyl alcohol (PVA) was obtained from Sigma Aldrich (St. Louis, MO, USA). CT26 cell line was procured from American Type Culture Collection (ATCC, USA) and fetal calf serum (FCS) was purchase from Biowest (USA). The cell line medium used was obtained from Hi media (Mumbai, India). 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT), propidium iodide, 4,6-diamidino-2 phenylindole (DAPI), trypsin-EDTA, and fluorescein isothiocyanate (FITC) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All the other chemicals and reagents used were purchased from Sigma-Aldrich (St. Louis, MO, USA).
6
Ginseng Compounds Inhibit Glioma and Colon Cancer
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WE, NSF, and SF of KRG were provided by the Korea Ginseng Cooperation (Daejeon, Korea). C6 glioma and CT-26 carcinoma cells were purchased from American Type Culture Collection (Manassas, VA, USA). Dulbecco's Modified Eagle's medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), streptomycin, penicillin, and L-glutamine were purchased from Gibco (Grand Island, NY, USA). Male Balb/c mice (age, 6–8 wk; weight, 17–21 g) were purchased from Orient Bio (Gyeonggi, Korea). Annexin V-FITC, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), and staurosporine were purchased from Sigma Chemical Co. (St Louis, MO, USA). TRI reagent was purchased from Molecular Research Center Inc. (Cincinnati, OH, USA). MuLV reverse transcriptase was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Polymerase chain reaction (PCR) premix and the primers used for semiquantitative reverse transcription PCR (RT-PCR) were obtained from Bioneer Inc. (Daejeon, Korea). The antibodies used in this study were purchased from Cell Signaling Technology (Beverly, MA, USA). The enhanced chemiluminescence system was purchased from AbFrontier (Seoul, Korea).
7
Regulation of Cell Proliferation by miR-30b-5p and PTEN
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Dulbecco’s modified Eagle’s medium (DMEM) was obtained from Hyclone (Logan, UT, USA); streptomycin, penicillin, PDGF, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI) were obtained from Sigma-Aldrich (Louis, MO, USA); fetal bovine serum (FBS) was obtained from (Louis, MO, USA); The pcDNA3.1-PTEN (PTEN), control empty vector (control), miR-30b-5p inhibitors, miR-30b-5p mimics as well as their negative controls (NC) were bought from RiboBio (Guangzhou, China); LipofectamineTM 2000 was obtained from Invitrogen (Carlsbad, CA, USA). TRIzol reagent was obtained from Vazyme (Nanjing, China); PrimeScript RT Master Mix and SYBR PrimeScript™ miRNA RT-PCR kit were obtained from Takara (Dalian, China); the primers were provided by BGI (Shenzhen, China); 5-bromo-2-Deoxyuridine (BrdU) solution was obtained from BD Pharmingen (San Diego, CA, USA); RIPA lysis buffer was obtained from Beyotime (Shanghai, China); BCA protein detection kit was obtained from Pierce Chemicals Co. Ltd. (Rockford, IL, USA); the antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA); enhanced chemiluminescence kit was obtained from Promega (Madison, WI, USA).
8
Cedrol and 5-FU Antitumor Effects
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Cedrol, purity more than 98% determined by GC analysis, was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), and 5-FU, dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), and RNase A were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies for various proteins, including p-p53, p21, p-Rb, PCNA, CDK4, cyclin D1, CDK2, cyclin A, cyclin B1, and FasL, were obtained from Santa Cruz Biotechnology (CA, USA). Primary antibodies against caspase-8, Bax, caspase-9 caspase-3, and β-actin were purchased from iReal Biotechnology (Hsinchu, Taiwan). Cedrol and 5-FU were dissolved in DMSO to a final concentration of 0.5%. An equal volume of DMSO was added as a control at a final concentration of 0.5% in the medium.
9
Semilicoisoflavone B Bioactivity Assay
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Semilicoisoflavone B (SFB) of ≥98% purity was purchased from ChemFaces (Wuhan, Hubei, China). SFB stock solution (100 mM) was prepared using dimethyl sulfoxide (DMSO, Sigma-Aldrich) and stored at −20°C. The DMSO concentration was less than 0.2% for each experiment. For the treatment with SFB, appropriate amounts of stock solution were administered to the medium to obtain the final experimental doses. Other chemical reagents used in the study including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), RNase A, DAPI dye, protease inhibitor cocktail, and phosphatase inhibitor cocktail were purchased from Sigma-Aldrich (St Louis, MO, USA). The primary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
10
Cloning and Purification of C. perfringens CPE
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The cpe open reading frame was PCR amplified from C. perfringens strain 8-6 with the primers P843 5′-ATGGATCCATGCTTAGTAACAATTTAAATCC-3′ and P979 5′-TGAATTCTTAAAATTTTTGAAATAATATTGAATAAGG-3′ adding BamHI-EcoRI and cloned into pGEX2T.
CPE was purified as previously described [56 (link)]. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI) were from Sigma (Paris, France). Cleavage by thrombin was performed with the Novagen thrombin kit (69022-3, Novagen Merck, Darmstadt, Germany) according to the manufacturer’s recommendations.
CPE was purified as previously described [56 (link)]. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI) were from Sigma (Paris, France). Cleavage by thrombin was performed with the Novagen thrombin kit (69022-3, Novagen Merck, Darmstadt, Germany) according to the manufacturer’s recommendations.
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