Dimethyl sulfoxide (dmso)
Manufactured by AppliChem
Sourced in Germany, United States, Spain
DMSO (Dimethyl Sulfoxide) is a clear, colorless, and odorless liquid that is commonly used as a solvent in laboratory settings. It has a high boiling point and is miscible with a wide range of organic and inorganic compounds, making it a versatile solvent for various applications in research and development.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (5 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Triton x 100, by AppliChem (4 mentions)
Triton x 100, by Merck Group (4 mentions)
Fetal calf serum (fcs), by PAN Biotech (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Fetal calf serum (fcs), by Merck Group (3 mentions)
Glutamine, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Cycloheximide (chx), by Merck Group (3 mentions)
Fluostar omega, by BMG Labtech (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Crystal violet, by AppliChem (3 mentions)
2 7 dichlorofluorescein diacetate dcf da, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Sartorius (3 mentions)
Tc20 automated cell counter, by Bio-Rad (2 mentions)
152 protocols using dimethyl sulfoxide (dmso)
1
Cryopreservation of PBMC Samples
2
5-Aza Effects on GSTP1 and TXNRD2
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Human LECs cell line SRA01/04 were treated with 10 μmol/L 5-Aza-2′-deoxycytidine (5-Aza, Sigma-Aldrich, USA), a DNMT inhibitor, for 3 days. 5-Aza was dissolved in dimethyl sulfoxide (DMSO, Applichem, Germany) and an equivalent amount of DMSO was used as a control treatment. To study the influence of 5-Aza on the expression of GSTP1 and TXNRD2, control and treated cells were collected and used for qPCR and western blot assays.
3
Investigating Aprepitant's Effects on HL60 Cells
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Human acute myeloid leukemia cell line HL60 cells were obtained from the Chinese Academy of Medical Sciences. Ara-C (Cat# C8040) was purchased from Solarbio and dissolved in PBS. Aprepitant (Cat# YZ-1041904) was purchased from Solarbio and dissolved in DMSO (Applichem, Cat# A3672) as the stock solution, while the content of DMSO in working solution is less than 0.1%. Z-VAD-FMK (Cat# HY-16658) was purchased from MCE. Necrostatin-1 (Cat# T1847) was purchased from TargetMol. Cells were cultured in RPMI medium 1640 (Gibco, Cat# 31800–022).
4
Evaluating ZKK-3 in Normoxic and Hypoxic Conditions
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The modified isothiourea derivative ZKK-3 (Fig. 1 ) was synthesized by Professor Zygmunt Kazimierczuk according to a previously described procedure (20 (link)). The compound was dissolved in dimethyl sulfoxide (DMSO; AppliChem GmbH, Darmstadt, Germany) and added to the culture medium at concentrations of 10, 25 and 50 µM. Control cultures were grown in standard conditions with DMSO but without ZKK-3 application (0 µM).
Cells were cultured under different gas mixtures with varying oxygen contents as follows: Normoxia (21% O2/5% CO2/74% N2 was applied for 24 h post-ZKK-3 treatment), anoxia (5% CO2/95% N2 was applied for 24 h post-ZKK-3 treatment); hypoxia (1% O2/5% CO2/94% N2 was applied for 24 h post-ZKK-3 treatment); HBO (97.5%O2/2.5% CO2 under pressure of 2 ATA was applied for 1 h post-ZKK-3 treatment, which was followed by 23 h of normoxia); hypoxia/hypoxia (double hypoxia; hypoxic gas (1% O2/5% CO2/94% N2) was applied for 24 h prior to ZKK-3 treatment, and then for an additional 24 h post-ZKK-3 treatment); and hypoxia/hyperbaric oxygen (hypoxia/HBO; hypoxia was applied for 24 h prior to ZKK-3 treatment, and HBO was applied post-ZKK-3 treatment). Anoxia and hypoxia experiments were performed in a Modular Incubator Chamber (MIC-101; Billups-Rothenberg, San Diego, CA, USA), whereas HBO experiments were conducted using a hyperbaric chamber (own design).
Cells were cultured under different gas mixtures with varying oxygen contents as follows: Normoxia (21% O2/5% CO2/74% N2 was applied for 24 h post-ZKK-3 treatment), anoxia (5% CO2/95% N2 was applied for 24 h post-ZKK-3 treatment); hypoxia (1% O2/5% CO2/94% N2 was applied for 24 h post-ZKK-3 treatment); HBO (97.5%O2/2.5% CO2 under pressure of 2 ATA was applied for 1 h post-ZKK-3 treatment, which was followed by 23 h of normoxia); hypoxia/hypoxia (double hypoxia; hypoxic gas (1% O2/5% CO2/94% N2) was applied for 24 h prior to ZKK-3 treatment, and then for an additional 24 h post-ZKK-3 treatment); and hypoxia/hyperbaric oxygen (hypoxia/HBO; hypoxia was applied for 24 h prior to ZKK-3 treatment, and HBO was applied post-ZKK-3 treatment). Anoxia and hypoxia experiments were performed in a Modular Incubator Chamber (MIC-101; Billups-Rothenberg, San Diego, CA, USA), whereas HBO experiments were conducted using a hyperbaric chamber (own design).
5
Epigenetic Modulation of Breast Cancer Cells
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MCF7, MDA-MB-231, MDA-MB-453, MDA-MB-468, BT-20, BT-474, ZR-75-1, HCC1937, MDA-MB-361, MDA-MB-157, and MCF12A cell lines were purchased from American Type Culture Collection (ATCC). MCF10A cell line was kindly provided by Elif Erson Bensan, (METU, Ankara, Turkey). All cell lines were grown as recommended by ATCC. AZA (5 μM, Sigma-Aldrich, A3656) dissolved in DMSO (Applichem, A1584.0100) was used to treat cells for 96 h, and equivalent amount of DMSO was used as control treatment. Media was changed at every 24th hour, and cells were harvested at 96th hour.
6
Combination Therapy for Myeloma
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The drugs used, Valproic acid (VPA; Depakine IV, Sanofi Aventis, Belgium) and Pioglitazone (PIO; Actos, Takeda, Japan), are commercially available. VPA was dissolved in sodium chloride (0.9%) as a 2M stock solution and a pill of PIO was reduced to powder and dissolved in DMSO (AppliChem, U.S.A.), at a concentration of 20mM. DMSO was tested alone for side effects.
After establishment of the MM mouse model, we studied the effect of single agents or a combination of them (VPA, PIO and VPA/PIO). Mice were divided in different groups and treated by intraperitoneal injection (IP) on a daily basis, for 2 weeks. Starting at day 10, mice were injected either with phosphate buffered saline pH 7.4, (PBS, Gibco, Life Technologies, UK), VPA and PIO or with combination of the last two.
Mocetinostat (MGCD0103, MethylGene Inc., Canada) and Vorinostat (Suberoylanilide Hydroxamic Acid; SAHA; Selleck Chemicals, U.S.A.), have been also used in-vitro, in combination to PIO. 2×105 MOLP8 cells were plated in culture flasks (Greiner, Belgium) and treated with MGCD0103 at 0.2μM, SAHA at 0.25μM or PIO at 30μM and combination of MGCD0103 and SAHA to PIO for 48 hours. BADGE (Bisphenol A Diglycidyl Ether, Sigma, Germany) has been used for its antagonist properties on PPARγ at 0.5μM concentration in co-treatment with PIO and HDACi.
After establishment of the MM mouse model, we studied the effect of single agents or a combination of them (VPA, PIO and VPA/PIO). Mice were divided in different groups and treated by intraperitoneal injection (IP) on a daily basis, for 2 weeks. Starting at day 10, mice were injected either with phosphate buffered saline pH 7.4, (PBS, Gibco, Life Technologies, UK), VPA and PIO or with combination of the last two.
Mocetinostat (MGCD0103, MethylGene Inc., Canada) and Vorinostat (Suberoylanilide Hydroxamic Acid; SAHA; Selleck Chemicals, U.S.A.), have been also used in-vitro, in combination to PIO. 2×105 MOLP8 cells were plated in culture flasks (Greiner, Belgium) and treated with MGCD0103 at 0.2μM, SAHA at 0.25μM or PIO at 30μM and combination of MGCD0103 and SAHA to PIO for 48 hours. BADGE (Bisphenol A Diglycidyl Ether, Sigma, Germany) has been used for its antagonist properties on PPARγ at 0.5μM concentration in co-treatment with PIO and HDACi.
7
Mass Spectrometry Sample Preparation
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Phosphate-buffered saline (PBS) was purchased from Wisent (Montreal, QC) and used without further purification. Dithiothreitol (DTT) was purchased from Merck (Whitehouse Station, NJ, USA). Sequencing grade-modified trypsin was purchased from Promega (Fitchburg, WI, USA). Iodoacetamide (IAA) and phosphatase inhibitor cocktail were purchased from Sigma (St. Louis, MO, USA). Complete protease inhibitor without ethylenediaminetetraacetic acid was from Roche Applied Science (Meylan, France). Dimethyl sulfoxide was purchased from Applichem (St. Louis, MO, USA). A BCA protein assay kit was purchased from Solarbio (Beijing, China). Benzonase was from Merck (Darmstadt, Germany). Titansphere Phos-TiO Kits were from GL Science (Japan). OASIS HLB sample extraction products were from Water Corporation (Milford, MA, USA). 3M Empore C8 disk was from 3M Bioanalytical Technologies (St. Paul, MN, USA). Water was double deionized. All other reagents used in the experiments were of mass spectrometry grade.
8
Isolation and Cryopreservation of PBMCs
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All blood samples were processed within 1 h of collection. PBMCs were isolated from heparin venous blood samples via density gradient centrifugation. In brief, samples were carefully layered on top of density medium (Biocoll, Biochrome, Germany). After centrifugation, PBMCs were harvested from the interphase, washed twice in phosphate-buffered saline (PBS) and taken up in RPMI-1640 + GlutaMax medium (Gibco, ThermoFisher Scientific, Germany) supplemented with 25% heat-inactivated fecal calf serum (FCS) (Biochrome, Germany) and 10% dimethylsulfoxide (Applichem GmbH, Germany) for cryopreservation. Cells were frozen at a concentration of 107 cells/ml at −80°C in a pre-cooled freezing container. After 24–48 h, cells were transferred to liquid nitrogen and stored at −196°C until analysis.
For thawing, the cryo vials were transferred to a water bath pre-warmed to 37°C. After 5 min, 107 cells were transferred into 10 ml of thawing medium (RPMI-1640 + GlutaMax containing 10% FCS at 37°C). Cells were then washed in medium, counted and prepared for phenotyping by flow cytometry or magnetic-activated cell separation (MACS Microbead Technology, Miltenyi Biotec, Germany) as described below.
For thawing, the cryo vials were transferred to a water bath pre-warmed to 37°C. After 5 min, 107 cells were transferred into 10 ml of thawing medium (RPMI-1640 + GlutaMax containing 10% FCS at 37°C). Cells were then washed in medium, counted and prepared for phenotyping by flow cytometry or magnetic-activated cell separation (MACS Microbead Technology, Miltenyi Biotec, Germany) as described below.
9
MTT Assay for Microglial Cell Viability
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Cell viability of Aβ1-42 oligomer-treated primary microglial cells and A1AT co-treated cells was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. Media was changed to serum-free media, and cells were treated with Aβ1-42 oligomers (2 μM) and A1AT (0.5, 2 and 4 mg/ml) for 24 hours. Culture medium was changed to fresh medium containing 0.5 mg/ml MTT and incubated for one hour at 37°C in a humidified atmosphere of 5% CO2. Medium was removed and cells were lysed with dimethyl sulfoxide (AppliChem, Darmstadt, Germany). The absorption was measured at 570 nm on a plate reader (ELISA-Reader Infinite™ 200series, Tecan, Crailsheim, Germany). Absorption values were normalized to untreated cells.
10
Hypoxia and Cell Signaling Assays
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Cells were cultured in DMEM, for HeLa and Huh7 cells, or DMEM F-12, for human bronchial smooth muscle (hBSM) cells (Lonza), containing 10% FCS and 100 U/ml penicillin/streptomycin (Biochrom). Cells were grown in a 37 °C incubator with 5% CO2. For hypoxic treatment, cells were exposed for 4–24 h to 1% O2, 94% N2 and 5% CO2 in an IN VIVO2 hypoxia workstation (Baker Ruskinn). When required, cells were treated for 4–24 h with CK1δ inhibitor D4476 (10 μΜ, Cayman Chemical) or kaempferol (50–100 μΜ, Sigma) using a 10 mM stock solution in dimethyl sulfoxide (Applichem). Transient transfections, reporter gene assays and chromatin immunoprecipitation were performed as previously described [6] (link).
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