Mouse hypothalamic neuronal GT1-7 cells (Sponsored by 10.13039/501100003347 Fudan University FBS (GIBCO) and 1% penicillin-streptomycin double-antibody solution (Biosharp), and maintained in a humidified environment at 37 °C with 5% CO2. The GT1-7 cells used in this study were all 3rd-10th generation. The mother liquor was prepared by dissolving 5-HT (MedChemExpress) in dimethyl sulfoxide (DMSO) (Beyotime), and the final concentration of DMSO in the medium was ≤0.1%.5-HT was prepared as follows: 5-HT was dissolved in DMSO to prepare a reserve solution with a final concentration of 2 mM and ultrasonicated in a cold water bath for 5 min and then stored at −80 °C.
Dimethyl sulfoxide (dmso)
Manufactured by Beyotime
Sourced in China, United States
DMSO is a colorless, odorless, and highly polar aprotic solvent used in various laboratory applications. It has a high boiling point and low viscosity, making it a versatile solvent for a wide range of chemical reactions and procedures. DMSO is known for its ability to dissolve a variety of organic and inorganic compounds, and it is commonly used as a solvent in spectroscopic and chromatographic analyses.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Beyotime (29 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Mtt solution, by Beyotime (13 mentions)
Mtt reagent, by Beyotime (9 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Microplate reader, by Bio-Rad (8 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Microplate reader, by Agilent Technologies (5 mentions)
Trypsin, by Thermo Fisher Scientific (5 mentions)
Bca protein assay kit, by Beyotime (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Microplate reader, by Molecular Devices (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Trypsin, by Beyotime (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Solarbio (3 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Beyotime (3 mentions)
Propidium iodide, by Beyotime (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
197 protocols using dimethyl sulfoxide (dmso)
1
Cell Culture and Neurotransmitter Preparation
2
Blue Light-Induced Photodamage in 661W Cells
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The 661W cells were seeded at 1 × 104 cells/well in 96-well plates and incubated for 24 h. The entire medium was then replaced with fresh medium containing 2% FBS. PKM2 inhibitor shikonin (Selleck, USA) or its solvent, dimethyl sulfoxide (DMSO; Beyotime, China), were added to the culture medium and incubated for 2 h. The final concentration of DMSO was not more than 0.1%. The cells were exposed to 6000 lux of blue light (wavelength~450 nm) for 3 h at 37°C to induce photodamage. The luminance was measured using a light meter (PP710, SanLiang, China). Calcein-acetoxymethyl ester (AM)/propidium iodide (PI) double staining, ROS assays, Cell Counting Kit-8 (CCK8) assays, and western blotting were performed after photodamage.
3
Atorvastatin Cytotoxicity Assessment Protocol
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Atorvastatin Calcium salt powder (C66H68 Ca F2N4O10, Molecular weight: 1155.363 g/mole) (Sigma Aldrich) is a hydrophobic substance, dissolved in organic solvents such as Dimethyl sulfoxide (DMSO), Dimethylformamide (DMF), and ethanol (U.S. National Library of Medicine, online document 2019 ; Product information 2018 ). These solvents themselves have shown a dose-dependent toxicity effect on MCF7 cells. If the maximum concentrations of ethanol and DMSO are 0.5% and DMF is 0.1%, this underlying effect is significantly reduced (Jamalzadeh et al. 2016 (link)). DMSO (Beyotime, Shanghai, China) has the lowest final concentration (less than 0.1% in DMEM medium), thus we used the DMSO as the atorvastatin solvent. After making the drug to a stock concentration of 100 nano mole per milliliter (nm/ml) in PBS (1.155 mg ATO + 100 μl DMSO + 9900 μl PBS), ATO diluted with DMEM- High glucose medium to the final concentrations of 5, 10, 15, 20, 30, 40, 50 (nm/ml) based on the M1V1 = M2V2 equation.
4
Atorvastatin Solubilization and Preparation
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Atorvastatin Calcium salt powder(C 66 H 68 Ca F 2 N 4 O 10 , Molecular weight:1155.363 g/mole) (Sigma Aldrich) is a hydrophobic substance, dissolved in organic solvents such as Dimethyl sulfoxide (DMSO), Dimethylformamide (DMF), and ethanol [28, 29] . These solvents themselves have shown a dose-dependent toxicity effect on MCF7 cells. If the maximum concentrations of ethanol and DMSO are 0.5% and DMF is 0.1%, this underlying effect is signi cantly reduced [30] (link). DMSO (Beyotime, Shanghai, China) has the lowest nal concentration (less than 0.1% in DMEM medium), thus we used the DMSO as the atorvastatin solvent. After making the drug to a stock concentration of 100 nano mole per milliliter (nm/ml) in PBS (1.155 mg ATO + 100 μl DMSO + 9900 μl PBS), ATO diluted with DMEM-High glucose medium to the nal concentrations of 5,10,15,20,30,40,50 (nm/ml) based on the M1V1=M2V2 equation.
5
Hypospermatogenesis Mouse Model Protocol
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All C57 male mice were supplied by the Southern Medical University Animal Center. This study was supported by the university's institutional animal care and use committee. The mouse testis tissues were collected at different postpartum times (P4-P10 weeks) for detecting PRPS2 expression. A mouse model of hypospermatogenesis was obtained as previously described.23 (link) Briefly, a total of 12 C57 male mice that were 8 weeks old were purchased and raised in the Southern Medical University Animal Center. All mice were randomly assigned to an experimental group or a negative control group. Those in the experimental group (n = 6) were treated with busulfan and dimethyl sulfoxide (DMSO; Biyuntian, Shanghai, China) solution (30 mg kg−1 body weight) by intraperitoneal injection. Mice in the control group (n = 6) were treated with the same volume of DMSO. Two weeks after treatment, the histopathological changes of the testis tissues were evaluated using hematoxylin–eosin (H and E; Yuanye, Shanghai, China) staining. Then, PRPS2 expression was quantified in the mice with hypospermatogenesis.
6
In Vivo Xenograft Tumor Model
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Five-week-old Balb/c nude mice were purchased from Vital River Laboratory Animal Technology Company (Beijing, China). Mice were randomly assigned to four groups (n = 5): MHCC-97H shC, MHCC-97H sh1, MHCC-97H shC + sorafenib, MHCC-97H sh1 + sorafenib. Equal amounts of MHCC-97H shC or MHCC-97H sh1 cells (1 × 107) were subcutaneously injected into the flank of each mouse and tumor volume determined. Tumor volume was calculated using the formula: volume = ab2/2 (a, the largest diameter; b, the perpendicular diameter). Drug administration was performed when the tumors reached a volume of ≈ 200 mm3. Mice received equal volume of sorafenib (15 mg/kg, #S7397, Selleck, Shanghai, China) or vehicle [DMSO (#196055, Biyuntian, Shanghai, China)] through intragastric administration every day for 3 weeks or less until the tumors reached 2000 mm3. Animals were sacrificed by cervical dislocation and the tumors excised and weighed. All animal procedures were performed in accordance with the guidelines of the Laboratory Animal Ethics Committee of Sun Yat-sen University.
7
Grifolin Preparation and Characteristics
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Grifolin was provided by the Kunming Institute of Botany; Chinese Academy of Sciences (Kunming, China) with the following structure: 2-trans, trans-farnesyl-5-methylresorcinol (purity, >99%; high performance liquid chromatography analysis). Grifolin was prepared at a concentration of 100 mmol/l stock solution in dimethyl sulfoxide (DMSO) and stored under −20°C so that the final concentration of DMSO was <0.1% in all assays it was used in. DMSO was provided by (Shanghai Biyuntian Bio-Technology Company, Ltd., Shanghai, China).
8
MTT Assay for Cell Viability
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3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) (Beyotime Institute of Biotechnology, Wuhan) was used to determine cell viability. MDA-MB-231 and MCF-7 cells (5000 cells/well) were seeded onto 96-well plate and then treated with different concentrations of 3-BP (0, 20, 40, 80, 160, 320 μmol·l-1) (Sigma-Aldrich, USA) when cells were attached completely. MTT solution (15 μl; 5 mg/ml in phosphate-buffered saline) were added into 96-well plate respectively at the 3-BP treatment of 24, 48, and 72 h. After 4 h, the solution was replaced with 150 μl dimethylsulfoxide (DMSO) (Beyotime Institute of Biotechnology, Wuhan) and 30 min later, cell viability was determined at a wavelength of 490 nm by a microplate reader (Synergy, BioTek, USA).
9
Cytotoxicity Evaluation of Lipid Nanoparticles
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SPI (≥92 %) and MCT (≥99 %) were obtained from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). DIO (≥98 %) was purchased from Nanjing Chunqiu Biological Engineering Co., Ltd. (Nanjing, China). Nile red and Nile blue (≥85 %) were bought from Sigma-Aldrich (Saint Louis, USA). Dulbecco's Modified Eagle's Medium (DMEM) and Hank's balanced salt solution (HBSS) were purchased from Shanghai BasalMedia Technologies Co., Ltd. (Shanghai, China). Fetal bovine serum (FBS) was obtained from Hyclone (Logan, USA). Penicillin-streptomycin solution, trypsin, and dimethyl sulfoxide (DMSO) were purchased from Beyotime Biotech. Inc. (Shanghai, China). Methyl Thiazolyl Tetrazolium (MTT) was from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). All other chemicals were of analytical reagent grade and were obtained from Beijing Chemical Works (Beijing, China).
10
Cytotoxicity Assay of J774A.1 Cells
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The J774A.1 cells were treated with imipramine (0, 25, 50, 75, 100 μmol/L), C2-Cer (0, 15, 30, 45, 60 μmol/L) and verapamil (0, 25, 50, 75, 100 μmol/L) for 24 h. It was followed by incubation of J774A.1 cells with MTT (Beyotime, Jiangsu, China) for 4 h. Furthermore, dimethylsulfoxide (DMSO) (Beyotime, Jiangsu, China) was added per well, and the absorbance was recorded by the microplate reader (BioTek, Winooski, VT, USA) at 490 nm.
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