The bivalent Smac mimetic birinapant was purchased from Medchemexpress (MCE, Medchemexpress CO. Ltd) and BV6 was purchased from Selleck (Blue Wood Chemical Co., Ltd.). Cells were treated with 10 µmol/L birinapant or BV6 as indicated, or with the equivalent amount of solvent dimethyl sulphoxide (DMSO, Beijing Dingguo Changsheng biotechnology CO. Ltd) 1 hour before irradiation. Irradiation with single doses of 2‐8 Gy was performed by a 137Cs γ‐source (Atomic Energy of Canada Limited, Gamma cell 40). The source‐target distance was 30 cm, and the dose rate was 0.99 Gy/min.
Dimethyl sulfoxide (dmso)
Manufactured by Dingguo
Sourced in China
Dimethyl sulfoxide (DMSO) is a colorless, odorless, and polar aprotic solvent. It has a high boiling point and is miscible with a wide range of organic and inorganic solvents. DMSO is commonly used as a solvent and vehicle for various chemical and biological applications in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin, by Dingguo (2 mentions)
Birinapant, by MedChemExpress (1 mentions)
Pentylenetetrazol, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Beyotime (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Mtt reagent, by Merck Group (1 mentions)
Crocin, by Yuanye Bio-Technology (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2 h tetrazolium bromide mtt, by Yuanye Bio-Technology (1 mentions)
Synergy 4 microplate reader, by Agilent Technologies (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Milli q ultrapure water system, by Merck Group (1 mentions)
Trypan blue, by Dingguo (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
96 well plate, by Corning (1 mentions)
Apigenin, by Maclin Biochemical Technology (1 mentions)
Baicalin, by Maclin Biochemical Technology (1 mentions)
Reduced glutathione, by Maclin Biochemical Technology (1 mentions)
11 protocols using dimethyl sulfoxide (dmso)
1
Birinapant and BV6 Sensitize Cells to Radiation
2
Neuroprotective effects of FPS-ZM1 in PTZ-induced seizures
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Drug: FPS-ZM1 (USA, Meck Millipore, Lot: 2908726), dissolved in a small amount of DMSO (Beijing Ding Guo chang Sheng Biotechnology Co., Ltd.) and then diluted with saline, In the end,the concentration of DMSO was 1%.
Animals: 130 clean grade Wistar male rats, 3-4 weeks age, weighing 50-60 g, were purchased from Shanghai Sleek Laboratory Animal Co., Ltd., License No.: SCXK (Shanghai) 2017-0005.
Reagents: Pentylenetetrazol (USA, Sigma, lot: MKCC1690), was con gured with saline the concentration of Pentylenetetrazol is 10 mg. ml-1. Western Blotting Assay Kit (KGP1201, Jiangsu Kaiji Biotechnology Co., Ltd.); photographic developer and xative (KGP116, Jiangsu Kaiji Biotech Development Co., Ltd.); HMGB1 ELISA Assay Kit (Shanghai Langton Biotechnology Co., Ltd., Lot:S20180318M)
Animals: 130 clean grade Wistar male rats, 3-4 weeks age, weighing 50-60 g, were purchased from Shanghai Sleek Laboratory Animal Co., Ltd., License No.: SCXK (Shanghai) 2017-0005.
Reagents: Pentylenetetrazol (USA, Sigma, lot: MKCC1690), was con gured with saline the concentration of Pentylenetetrazol is 10 mg. ml-1. Western Blotting Assay Kit (KGP1201, Jiangsu Kaiji Biotechnology Co., Ltd.); photographic developer and xative (KGP116, Jiangsu Kaiji Biotech Development Co., Ltd.); HMGB1 ELISA Assay Kit (Shanghai Langton Biotechnology Co., Ltd., Lot:S20180318M)
3
Isolation and Expansion of Rat Bone Marrow-Derived Mesenchymal Stem Cells
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BMSCs were obtained from the bone marrow of 2-week-old male Sprague-Dawley (SD) rats (n = 16, obtained from Chongqing Medical University Animal Care Centre, Chongqing, China) as described previously [22 (link)]. In detail, bone marrow was rinsed from femurs and tibia using culture medium that contained Dulbecco's modified Eagle's medium (DMEM)/F-12 (GIBCO, Grand Island, NY, USA), 10% fetal bovine serum (FBS) (AusGeneX, Brisbane, Australia), and 1% penicillin and streptomycin. BMSCs were obtained by whole bone marrow differential adherence method. In the first 24, 48, and 72 hours, unadhered cells could be effectively exchanged using total medium exchange. Thereafter, the medium was changed every 3 days. When the density of cells reached 95%, adherent BMSCs were digested with trypsin (GIBCO) and passaged on. Third-generation BMSCs were stored in a 90% FBS and 10% dimethyl sulfoxide (DMSO) (Dingguo Biotech, Beijing, China) mixture in liquid nitrogen. If necessary, BMSCs were resuscitated by rapid thawing in a 37°C water bath, plated in culture medium, and cultured at 37°C in humid air with 5% CO2. Third-generation BMSCs were used for all experiments [23 (link)]. BMSCs were maintained in 37°C and 5% CO2 incubators (Thermo Fisher Scientific, Waltham, MA, USA).
4
Ginger Oleoresin Cytotoxicity and Radioprotection
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Ginger oleoresin was presented by Tianjin University of Commerce and diluted in the equal volume of dimethyl sulfoxide (DMSO) (Dingguo, BJ, China) to prepare the stock solution. Then, the stock solution was further diluted to the required concentration using cell culture medium. For cytotoxicity determination of ginger oleoresin, hMSCs were exposed to 10−3, 10−4, 10−5, and 10−6 g/mL ginger oleoresin and cultured for 24 h, 48 h, and 72 h. For radioprotective effect determination, hMSCs were pretreated with 10−4 g/mL ginger oleoresin and cultured for 2 h followed by exposure to γ-rays at the Irradiation Center. After irradiation, the cells were subsequently incubated for 24 h. 137Cs was used as the irradiation source (AECL, Canada). hMSCs cells (±ginger oleoresin in medium) were irradiated at a dose of 4 Gy. DMSO was used as a vehicle control. hMSCs + ginger oleoresin without γ-ray irradiation were also studied.
5
Cell Viability Assay for Mesenchymal Stem Cells
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Cell viability was assessed by MTT (Beyotime, Shanghai, China) assay. MTT assay was performed at 0, 24, 48, and 72 hours after transfection. Bone marrow mesenchymal stem cells were seeded in 96-well plates at a density of 1 × 104 cells/well (100 μL/well). Then 20 μL MTT (5 mg/mL) was added in each well, and maintained for 4 hours. The medium was removed, and 200 μL dimethyl sulfoxide (Dingguo, Beijing, China) was added in each well, followed by 10 minutes of oscillation. Optical density at 490 nm was measured using an enzyme linked immunosorbent assay. Cell survival rates were calculated by the ratio of absorbance between the transfection group and the untransfected group.
6
MTT Assay for Cell Viability
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Cell viability was determined using an MTT assay. Briefly, the cells (1×104 cells/well) were plated for 24 h in 96-well plates in 200 µl complete medium and exposed to different concentrations of inhibitors for various durations. Each treatment was repeated in six separate wells. The cells were incubated at 37°C with 5% CO2, and MTT reagent (20 µl, 5 mg/ml; Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBS) was added to each well and incubated for 4 h. The formazan crystals were dissolved in 150 µl dimethyl sulfoxide (Beijing Dingguo Changsheng Biotechnology Co., Ltd., Beijing, China) and the absorbance was recorded at a wavelength of 490 nm using a Microplate Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA). Cell viability was calculated as follows: Cell viability (%) = absorbanceexperiment/absorbancecontrol × 100.
7
Crocin Cytotoxicity Evaluation in SW480 Cells
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SW480 cells were seeded into 96-well plates at a density of 8 × 103 cells/well. After incubation at 37 C overnight, cells were treated with 0.4, 0.8, 2, 3 and 6 mM of crocin (catalog number: B21336; Shanghai Yuanye Biological Technology Co., Ltd., Shanghai, China) for 24 h at 37 C. Thereafter, 5 μl of 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) (Shanghai Yuanye Biological Technology Co., Ltd., Shanghai, China) was added to each well, to obtain a final concentration of 5 mg/ml MTT. The plates were then incubated at 37 C for a further 4 h, at which point the supernatant was aspirated, 100 μl of dimethyl sulfoxide (catalog number: DH105-2; Beijing Dingguo Changsheng Biotechnology Co., Ltd., Beijing, China) was added to each well, and finally the absorbance of the plates was measured at 490 nm using a Synergy™ 4 Microplate Reader (Biotek, Winooski, Vermont, United States).
8
Cell Culture Protocol for ACS Extraction
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ACSs were extracted with ethanol, dissolved in phosphate buffered saline (PBS) (pH 7.8; China), and diluted in cell culture medium for preparation. Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), 2 mmol/L l -glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin were purchased from Gibco Life Technologies (Grand Island, USA). Giemsa and hematoxylin-eosin (HE) stains were purchased from Ameresco (USA). 5-Fluorouracil (5-Fu), l -Dopa, bovine serum albumin, and phenylmethanesulfonyl fluoride (PMSF) were purchased from BBI (USA). Acridine orange (AO), ethidium bromide (EB), dimethyl sulfoxide, trypan blue, trypsin, and ethylenediaminetetraacetic acid were purchased from Beijing Dingguo Changsheng Biotech Co., LTD. (China). Triton X-100 and Coomassie Brilliant Blue G250 were obtained from Shanghai Sangon Biological (China). Six orifice plates and tissue culture flasks were purchased from NUNC Company (Denmark). Other chemicals were purchased from local companies. Solutions were prepared using a Milli-Q ultrapure water system (Millipore Corp.).
9
Cell Viability Assessment by MTT Assay
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Cell viability was determined by standard 3-(4,5-dimethylthiazol-2-yl)-2,4-diphenyl-tetrazolium bromide (MTT, Sigma-Aldrich) assay. After transfection, cells were implanted at a density of 2 × 104 cells per well in 96-well plates (Corning) for 48 h or overnight with Gemcitabine treatment for 48 h. Briefly, 5 mg mL−1 MTT (Sigma) was added and incubated at 37 °C for another 4 h; thereafter, the medium was replaced and the formazan crystals were dissolved in 150 μL of dimethyl sulfoxide (DMSO, Dingguo, Beijing, China). The optical density (OD) was determined with a Thermomax microplate reader (Bio-Tek EL, USA) at 490 nm wavelength. All experiments were performed in triplicate and repeated at least three times.
10
Green Tea-Mediated Amyloid Inhibition
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Yunwu green tea is produced in Zhejiang Province, China. HAuCl4•3H2O (CAS: 16961-25-4, 99.9%), and hesperidin (CAS: 520-33-2, 98%) were purchased from Aladdin (Shanghai, China). Amyloid β protein fragment 1–42 (Aβ42, 107761-42-2, 95%), EGCG (CAS: 989-51-5, 98%), quercetin (CAS: 117-39-5), apigenin (CAS: 520-36-5), reduced glutathione (CAS: 70-18-8, 99%), baicalin (21967-41-9, 98%), thioflavin T (CAS: 2390-54-7, 97%) and hexafluoroisopropanol (CAS: 920-66-1, 99.5%) were purchased from Macklin (Shanghai, China). Dimethyl sulfoxide (DMSO) and phosphate-buffered solution (PBS, 10 mM, pH 7.2–7.4) were obtained from Dingguo Changsheng Biotechnology Co., Ltd (Beijing, China) and Genview (Beijing, China), respectively. Isopropanol (CAS: 67-63-0, 99.9%) was purchased from Concord Technology Co., Ltd (Tianjin, China). 18.2 MΩ/cm2 water was obtained in-house using a UPH-II-20T ultrapure water manufacturing system (Sichuan, China).
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