Harmol was purchased from Tokyo Chemical Industry. Doxycycline was purchased from MedChemExpress. Dorsomorphin dihydrochloride was purchased from Targetmol. G418 sulfate was purchased from Amresco. Dopamine hydrochloride, homovanillic acid, 3, 4-dihydroxyphenylacetic acid, tri-sodium citrate dihydrate, citric acid, chloroquine, anti-Flag M2, and anti-α-synuclein antibodies were purchased from Sigma-Aldrich. anti-TFEB antibody was purchased from Proteintech. Anti-tyrosine hydroxylase, anti-phospho-mTOR (Ser2448), anti-mTOR, anti-phospho-ULK1 (Ser757), anti-phospho-ULK1 (Ser317), anti-phospho-ULK1 (Ser555), anti-ULK1, anti-phospho-AMPKa (Thr172), anti-AMPKa, anti-LAMP1, anti-SQSTM1/p62, anti-TFEB, and anti-CTSD/cathepsin D antibodies were purchased from Cell Signaling Technology. Anti-phospho-α-synuclein and anti-LC3B antibodies were purchased from Abcam. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was purchased from Beyotime Biotechnology.
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide
Manufactured by Beyotime
Sourced in China
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide is a chemical compound used in laboratory research. It is a tetrazolium salt that undergoes reduction in the presence of metabolically active cells, resulting in the formation of a colored formazan product. This reaction is commonly used as an indicator of cell viability and proliferation in various cell-based assays.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Beyotime (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Bcl 2, by Proteintech (2 mentions)
Homovanillic acid, by Merck Group (1 mentions)
Doxycycline, by MedChemExpress (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Trisodium citrate dihydrate, by Merck Group (1 mentions)
Anti tfeb antibody, by Proteintech (1 mentions)
Anti lamp1, by Cell Signaling Technology (1 mentions)
Anti tyrosine hydroxylase, by Cell Signaling Technology (1 mentions)
G418 sulfate, by Avantor (1 mentions)
Anti sqstm1 p62, by Cell Signaling Technology (1 mentions)
Anti lc3b antibody, by Abcam (1 mentions)
Anti tfeb, by Cell Signaling Technology (1 mentions)
3 4 dihydroxyphenylacetic acid, by Merck Group (1 mentions)
Citric acid, by Merck Group (1 mentions)
Dopamine hydrochloride, by Merck Group (1 mentions)
Anti flag m2, by Merck Group (1 mentions)
Annexin 5 fitc pi double staining apoptosis detection kit, by Beyotime (1 mentions)
Shp 2, by Proteintech (1 mentions)
10 protocols using 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide
1
Biochemical Assays for Autophagy
2
Apoptosis and ROS Regulation in Cancer Cells
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PSN-A (Figure 1 A) was purchased from EXTRASYNTHESE (Genay, France). Dulbecco's Modified Eagle's Medium (DMEM) and fatal bovine serum (FBS) were purchased from Gibco (Eggenstein, Germany) while Penicillin/ Streptomycin antibiotics was obtained from Solarbio co., Ltd. (Beijing, China). Annexin V-FITC/PI double staining apoptosis detection kit, Reactive Oxygen Species (ROS) assay kit, mitochondrial membrane potential (MMP) assay kit, Crystal violet, 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), dimethyl sulfoxide (DMSO), Fluo-3AM and doxorubicin were purchased from Beyotime Institute of Biotechnology (Haimen, Jiangsu, China). The primary antibodies for cleaved caspases-3, cleaved PARP, p-STAT3 (Tyr705), STAT3 and SHP-2 were purchased from Cell Signaling Technology (Beverly, MA) while primary antibodies for Bax, Bcl-2, SHP-2, PTEN and β-actin were purchased from Proteintech (Wuhan, China). The primary antibodies for p-JAK2 and JAK2 were purchased from abcam (Cambridge, MA). Horseradish peroxidase (HRP)-conjugated secondary antibodies (goat anti-rabbit, goat anti-mouse) were obtained from Sigma.
3
Synergistic Antioxidant Bioactive Compounds
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6-shogaol (98% purity, analytical-grade reagent) was purchased from Aladdin Industrial Corporation (Shanghai, China). Curcumin was obtained from J&K Scientific Co. Ltd. (Beijing, China). Ginger extractive (10% gingerol) was purchased from Nanjing Zelang Biological Technology Co. Ltd. (Nanjing, China). mPEG2K-LA was synthesized in our laboratory [32 (link)]. (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypsin were provided by the Beyotime Institute of Biotechnology (Jiangsu, China). Diagnostic kits for assaying serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and the glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD) and the levels of malondialdehyde (MDA) were purchased from Nanjing Jian Cheng Bioengineering institute (Nanjing, China). Chromatographically pure methanol and acetonitrile were procured from Sinopharm Chemical Reagent Technology Co. Ltd. (Shanghai, China).
4
Pitavastatin's Effect on Liver Cell Viability
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The Huh-7 cells and SMMC7721 cells were split into 96-well dishes at 5,000 cells/well and treated with the indicated dosage of pitavastatin for 48 hours or 5 µM pitavastatin for 1, 2, 4, 6 days respectively. The cells were incubated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (Beyotime, Haimen, People’s Republic of China) and formed formazan in the liver cells. Formazan was dissolved in dimethyl sulfoxide (DMSO; Beyotime), and the absorbance was measured at the wavelength of 570 nm. The cells treated with DMSO were used as a control group. The relative cell number of each group was calculated as pitavastatin-treated group/cell number in the DMSO-treated group.
5
Dihydroartemisinin-Nile Red Nanoparticle Synthesis
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Dihydroartemisinin (DHA) and Nile Red were bought from Aladdin (Shanghai, China). 2-nitroimidazole (2-MI) and indocyanine green (ICG) were purchased from J&K Scientific (Beijing, China). Zinc nitrate and bovine serum albumin (BSA) were purchased from Sigma–Aldrich (Merck Life Science, Shanghai, China). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 4, 6-diamidino-2-phenylindole (DAPI), and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were bought from Beyotime Biotechnology (Shanghai, China). DMEM medium and fetal bovine serum (FBS) were bought from Shanghai Titan Scientific Co., Ltd., (Shanghai, China). CAAT/enhancer binding protein (C/EBP) homologous protein (CHOP), Bcl-2, and β-actin antibodies were bought from Cell Signaling Technology, Inc., (CST, Boston, MA, USA).
Cells were kindly provided by Pricella Life Science&Technology Co., Ltd., (Wuhan, China).
Cells were kindly provided by Pricella Life Science&Technology Co., Ltd., (Wuhan, China).
6
Apoptosis and Oxidative Stress Assays
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Gen and Rev were provided by Xi’an QingYue Biotechnology Co. Ltd. (China) and Sigma Chemical Co. (St. Louis, MO, USA), respectively. As2O3 was acquired from Harbin YI-DA Pharmaceutical Limited Company. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), cell-penetrating lipophilic cationic fluorochrome JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazole-carbocyanide iodine), the Total Superoxide Dismutase Assay Kit with 2-(4-iodophenyl)- 3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-1), and Annexin V-FITC Apoptosis Detection Kit were bought from Beyotime Institute of Biotechnology (China) and stored at −20°C in the dark. The 2′,7′-dichlorofluorescein diacetate (DCFH-DA) was provided by Molecular Probes (Eugene, OR, USA). The TUNEL detection kit was purchased from Roche (Cell Death Detection Kit; Roche Biochemicals; Mannheim, Germany). LC3A/B monoclonal antibody was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).
7
Santamarine Cytotoxicity and Apoptosis
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Santamarine (purity > 95%) was purchased from BioBioPha. Co., Ltd. (Kunming, China). Hoechst 33258, Propidium Iodide (PI), Calcein acetoxymethylester (Calcein-AM), Dimethyl sulfoxide (DMSO), [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Crystal violet staining solution and N-acetyl L-cysteine (NAC) were purchased from Beyotime Institute of Biotechnology (Nanjing, China). IKK-16 (I-кB Kinase Inhibitor) and S31.201 (STAT3 inhibitor) was purchased from Selleckchem. (Shanghai, China). Human TNF-α was purchased from Sino Biological technology (Beijing, China). Dulbecco's Modified Eagle's Medium (DMEM) was purchased from GIBCO (Shanghai, China). Penicillin and Streptomycin was purchased from Solarbio co., Ltd. (Beijing, China). Fetal Bovine Serum (FBS) was purchased from Tissue culture biologicals (TCB, America). Antibodies specific to Bax, bad, Bcl2, cytochrome c, caspase 3, -8, Bid, PARP, TrxR and p50 were purchased from Protein tech. (Wuhan, China). Survivin, p65, histone H3 and IкB-α was purchased from Beyotime (Nanjing, China) while p-IкB-α was purchased by RuiYing, (Nanjing, China). SHP2, Src, p-Src, STAT3, p-STAT3 were purchased from cell signaling technology.
8
Cell Viability Assay with Irradiation
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Ishikawa or HEC-1B cells were treated according to the corresponding protocols and then seeded in 96-well plates. Twenty-four hours later, cells were treated with 4 Gy irradiation at a dose of 0.5 Gy/min (Varian2300EX, Varian, USA). Seven-two hours later, cells in each well were incubated with 10 µl 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (5 mg/ml; Beyotime Biotechnology, China) for 4 h and 100 µl Fromazan for another 4 h. Then, the absorbance of each well at 570 nm was examined using a microplate reader.
9
Purified ANTX-a Assay Protocol
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Purified ANTX-a (CAS No. 64285-06-9, C10H15NO, MW = 165.23) was purchased from Sigma (St. Louis, MO, United States). Lymphoprep was purchased from Huadong Pharmaceutical (Zhejiang, China). Fetal calf serum (FCS) and RPMI-1640 medium were acquired from Hangzhou Key Shengwu (Hangzhou, China). Malonaldehyde (MDA), SOD, glutathione (GSH), CAT, glutathione-s-transferase (GST), glutathione reductase (GR), and glutathione peroxidase (GPx) assay kits were all purchased from Nanjing Jiancheng Bioengineering, Inc. (Jiangsu, China).
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, propidium iodide (PI), and rhodamine 123 assay kits were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Other reagents used were purchased from commercial sources unless otherwise specified.
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, propidium iodide (PI), and rhodamine 123 assay kits were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Other reagents used were purchased from commercial sources unless otherwise specified.
10
Extraction and Quantification of Algal Carotenoids
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The solvents used for the preparation of the algal carotenoid extract, including acetonitrile, methanol (MeOH), methylene chloride (CH2Cl2), ethanol, and n-hexane, were purchased from Merck Co. (Darmstadt, Germany). Distilled deionized water (H2O) was prepared with the Ultrapure water purification system (Lotun Co., Ltd, Taipei, Taiwan). Potassium hydroxide (KOH) was purchased from Merck Co. All-trans forms of lutein (MW 569) and zeaxanthin (MW 569), glutamine, penicillin, streptomycin, DMSO, Griess reagent (1% sulfanilamide, 0.1% N-(1-naphthyl) ethylenediamine dihydrochloride, and 2.5% phosphoric acid), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, EDTA, leupeptin, phenylmethanesulfonyl fluoride, phosphate-buffered saline (PBS), sodium chloride (NaCl), sodium nitrite (NaNO2), Triton X-100, Tris, AG490, and STAT-3 inhibitor were purchased from Beyotime Inc. (Shanghai, China). RPMI1640, fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Hyclone (Logan, UT, USA). Enzyme-linked immunosor-bent assay (ELISA) kits for the measurement of IL-6 were purchased from eBioscience (San Diego, CA, USA). The inhibitors of JAK2 (AG490) and STAT-3 were obtained from Cayman Chemical (Michigan, USA).
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