A molecule library containing 2320 compounds (SPECTRUM 2320) was obtained from MicroSource Discovery Systems. Compounds were pre-dissolved to 10 mM in DMSO (Biosolve) and final DMSO content was kept constant at a well-tolerated 0.2% (v/v) for all in vitro experiments. Celastrol was derived from this library and performed equally well compared to celastrol obtained from another source (Sigma Aldrich).
Dimethyl sulfoxide (dmso)
Manufactured by Biosolve
Sourced in Netherlands
DMSO (Dimethyl Sulfoxide) is a colorless, odorless, and polar aprotic solvent. It has a high boiling point and is miscible with water and many organic solvents. DMSO is commonly used as a solvent and cryoprotectant in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Acetonitrile, by Biosolve (3 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lysis reagent, by Promega (2 mentions)
Hexane, by Biosolve (2 mentions)
Ultra pure ethanol, by Merck Group (2 mentions)
Dulbecco s modification of eagle s medium high glucose dmem without phenol red, by Merck Group (2 mentions)
Glass fiber filters mg 227 1 60, by Sartorius (2 mentions)
Luciferine reagent, by Promega (2 mentions)
17β estradiol, by Merck Group (2 mentions)
Sodium pyruvate, by Merck Group (2 mentions)
Acetone, by Biosolve (2 mentions)
N hydroxysuccinimide nhs, by Thermo Fisher Scientific (1 mentions)
N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride edc, by Merck Group (1 mentions)
175 cm2 tissue culture flasks, by Corning (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Nahco3, by Merck Group (1 mentions)
17 protocols using dimethyl sulfoxide (dmso)
1
Screening Compound Library for Bioactive Molecules
2
Functionalization of Mesoporous Silica Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Modification of the MSNs with PEG4-N3 was performed (as described in [23 (link)]) to inhibit interactions between the peptide and the MSN, thereby protecting the conformation of the peptide. In short, stirring 291.3 mg 15-Azido-4,7,10,13-tetraoxa-pentadecanoic acid (N3-PEG4-COOH, 1 mmol, >98%, Iris Biotech, Marktredwitz, Germany) with 140 mg N-hydroxysuccinimide (NHS, 1.2 mmol, ≥98%, Alfa Aesar, Haverhill, MA, USA) in 1 mL dimethylsulfoxide (DMSO, Biosolve, Valkenswaard, The Netherlands) containing 400 mg N-(3-(dimethylamino)propyl)-N’-ethylcarbodiimide hydrochloride (EDC, 2.1 mmol, Sigma-Aldrich) and 160 mg hydroxybenzotriazole (HOBt, 1.2 mmol, Biosolve) for 30 min under N2 gas atmosphere in an ice bath resulted in the N-hydroxysuccinimide ester of N3-PEG4-COOH (NHS-1). Next, 100 mg of MSN-NH2 in 4 mL DMSO was added to the NHS-1 reaction mixture and stirred under N2 gas atmosphere for 72 h at room temperature, after which the mixture was filtered and washed with deionized water and ethanol (96%) to obtain MSN-PEG4-N3.
3
Cryopreservation of PC-3 Prostate Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human prostate adenocarcinoma cell line PC-3 was cultured in Dulbecco’s modified Eagle’s medium (DMEM)–F12 (Ham’s) growth medium (Gibco Fisher Scientific, Landsmeer, Netherlands), supplemented with 10% fetal bovine serum (FBS) (Gibco Fisher Scientific), 1 mM sodium pyruvate (Gibco Fisher Scientific), 1.5 g/L NaHCO3 (Merck, Schiphol-Rijk, Netherlands), 0.1 mM nonessential amino acids (Gibco Fisher Scientific), and 50 µg/mL gentamycin (Sigma Aldrich, Zwijndrecht, Netherlands). PC-3 cells were grown in 175-cm2 tissue culture flasks (Corning, Amsterdam, Netherlands) in culture medium as described above. Before reaching maximal density, cells were washed with 1× phosphate-buffered saline (PBS) (Sigma Aldrich) and Trypsinized with 1× Trypsin (Gibco Fisher Scientific). Subsequently, medium was added and cells were pelleted by centrifugation and resuspended in FBS with 10% DMSO (Biosolve B.V., Valkenswaard, Netherlands) and stored in aliquots at −150 °C.
4
NMR Characterization of Ubiquitin Conformations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freeze-dried ubiquitin and diubiquitin samples were dissolved in 5% DMSO (Biosolve) in Milli-Q® water and then redissolved in NMR buffer containing 20 mM NaPO4 pH 6.8 and 10% D2O. Then, samples were taken in 15 ml 3.5 kDa Millipore spin filter tubes and spun-washed with three volumes of NMR buffer until DMSO was almost completely removed (LC/MS analysis). Concentrated samples were diluted to 500 μL with NMR buffer and the final concentration was determined using BCA assay using ubiquitin as standard. The pH was carefully measured using a Mettler TOLEDO pH probe.
All NMR studies were carried out on a Bruker 900 MHz spectrometer with a TCI cryoprobe, at 298 K (25°C). [1H, 15N] HSQC-spectra were acquired, processed and calibrated using standard methods. Chemical Shift Perturbations (CSPs) were calculated by comparing the [1H, 15N] HSQC spectra of mono Ub with that of each of the diUb molecules/ The CSP was calculated according to the following formula
where ΔδH and ΔδN are the chemical shift differences for 1H and 15N, respectively.
The spectra of K6 diUb indicated two different co-existing conformations. An “open conformation” was assigned based on similarity with the mono-Ub spectrum.
All NMR studies were carried out on a Bruker 900 MHz spectrometer with a TCI cryoprobe, at 298 K (25°C). [1H, 15N] HSQC-spectra were acquired, processed and calibrated using standard methods. Chemical Shift Perturbations (CSPs) were calculated by comparing the [1H, 15N] HSQC spectra of mono Ub with that of each of the diUb molecules/ The CSP was calculated according to the following formula
where ΔδH and ΔδN are the chemical shift differences for 1H and 15N, respectively.
The spectra of K6 diUb indicated two different co-existing conformations. An “open conformation” was assigned based on similarity with the mono-Ub spectrum.
5
Cryopreservation of mIMCD3 Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All mIMCD3 cell lines were cultured in 175 cm2 culture flasks at 37 °C in an environment of 5% CO2 in DMEM/F12 (Ham’s) culture medium (D8062, Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS, Gibco Fisher Scientific, Landsmeer, Netherlands), Glutamax, and penicillin/streptomycin. Before maximal cell density was reached, cells were washed with 1× phosphate-buffered saline (PBS) (Sigma-Aldrich) and trypsinized with 1× trypsin (Gibco Fisher Scientific). Medium was subsequently added and cells were pelleted by centrifugation for 5 min at 1500 rpm. The cell pellet was resuspended in FBS with 10% DMSO (Biosolve B.V., Valkenswaard, Netherlands) and frozen in aliquots to −150 °C.
6
Analytical Workflow for Pharmaceuticals
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All solvents were of U(H)PLC grade. Merck KGaA supplied MeCN, and MeOH was purchased from Reuss-Chemie AG. Purified water was obtained from a Milli-Q integral water purification system.
Scharlau supplied DMSO, and formic acid was from BioSolve. Antipyrine and BSA were purchased from Sigma-Aldrich and Antipyrine-D3 from HPC Standards GmbH. Protopine HCl was purchased from
Extrasynthese SAS, and verapamil HCl from Sigma-Aldrich. Citalopram HBr, diazepam, and diazepam-D5 were purchased from Lipomed AG, and citalopram-D4 HBr was obtained from CDN Isotopes.
Scharlau supplied DMSO, and formic acid was from BioSolve. Antipyrine and BSA were purchased from Sigma-Aldrich and Antipyrine-D3 from HPC Standards GmbH. Protopine HCl was purchased from
Extrasynthese SAS, and verapamil HCl from Sigma-Aldrich. Citalopram HBr, diazepam, and diazepam-D5 were purchased from Lipomed AG, and citalopram-D4 HBr was obtained from CDN Isotopes.
7
Spin-Labeling of Amyloid-Beta Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The A β40 peptide as well as two cysteine-A β40 variants: [cys26] –A β40 and [cys1] –A β40, differing in the position of the spin label, were purchased from AnaSpec (purity > 95%), the solvent DMSO was purchased from Biosolve (purity 99.8%), the spin probe MTSSL [1-Oxyl-2,2,5,5-Tetramethyl- Δ-Pyrroline-3-Methyl] Methanethiosulfonate was purchased from Toronto Research Chemicals Inc. (Brisbane Rd., North York, Ontario, Canada, M3J 2J8). Spin labeling was performed and the purified spin-labeled A β was analyzed by liquid chromatography and liquid chromatography/mass spectrometry as described previously [42 (link)]. The spin-labeled construct thus obtained is referred to as SL1-A β or SL26-A β. The peptide was lyophilized and stored in the freezer (–20°C) until used.
8
Microbial Metabolite Extraction Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Interaction zones (Figure 1 A) from three independent Petri dishes were excised and pooled together in a flask with one volume of ethyl acetate (99.8%, Biosolve, Dieuze, France). After 90 min of agitation (120 rpm) at room temperature, ethyl acetate extracts were recovered by filtering on a paper filter of 100 µm and were vacuum-dried at room temperature. Dry extracts were resuspended at a concentration of 10 mg/mL either in DMSO (extra dry, 99.8%, Biosolve) or in methanol (HPLC quality, 99.9%, Carlo Erba, Val de Reuil, France). For fractioning experiments, 80 Petri dishes were pooled to obtain the final extract.
9
Elucidating Structure-Retention Relationships
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To elucidate the effects of molecular structure on both 1tR and 2tR, a model compound (MC) mixture was prepared and analyzed by means of GPC‐HPLC‐UV/VIS. The mixture contains 2000 mg/mL of 18 different model components diluted in dimethylsulfoxide (DMSO, Biosolve, 99.9+%). The molecular structures of all model compounds, categorized into 3 groups, are presented in Figure 1 . The first group are phenolics (blue in Figure 1 ); phenol (Ph, Chem‐Lab, 99+%), resorcinol (R, Chem‐Lab, 99+%), p‐cresol (PC, Sigma‐Aldrich, 99+%), guaiacol (G, Sigma‐Aldrich, 99+%), vanillin (V, Sigma‐Aldrich, 99 %) and ethyl vanillin (EV, Sigma‐Aldrich, 98.5 %). The second group are eugenolics (green in Figure 1 ); eugenol (EUG, Sigma‐Aldrich, 99+%), coniferyl alcohol (CA, Alfa Aesar, 98 %), sinapyl alcohol (SA, Sigma‐Aldrich, 80 %), trans‐ferulic acid (TFA, Sigma‐Aldrich, 99 %) and 3‐(3‐ethoxy‐4‐methoxyphenyl)‐1‐propene (EMP, ABCR, 95 %). The third and final group are dimerics (violet in Figure 1 ); 4‐phenoxyphenol (4PP, Sigma‐Aldrich, 99 %), benzyl‐phenylether (BPE, Alfa Aesar, 97 %), 1‐phenyl‐2‐phenoxyethanol (PPE, AmBeed, 97+%), guaiacylglycerol‐beta‐guaiacylether (GG, Sigma‐Aldrich, 97+%), 6,6′‐dihydroxy‐5,5′‐dimethoxybiphenyl‐3,3′‐dicarbaldehyde (DDC, ABCR, 95 %), 1‐(3,4‐dimethoxy‐phenyl)‐2‐(2‐methoxy‐phenoxy)‐propane‐1,3‐diol (DMD, ABCR, 97 %) and combretastatin A4 (COM, Sigma‐Aldrich, 98+%).
10
In vitro metabolism of synthetic cathinones
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acetonitrile (LC-MS grade) and DMSO were purchased from Biosolve (Valkenswaard, The Netherlands). Formic acid (LC-MS grade) was obtained from Merck (Zwijndrecht, The Netherlands). Ultra-pure water was obtained from a Milli-Q Plus purification system (Millipore, Amsterdam, The Netherlands). Magnesium chloride, NADPH, glucose-6-phosphate, and glucose-6-phosphate dehydrogenase were purchased from Sigma (Zwijndrecht, The Netherlands). Pooled male rat (Sprague-Dawley) liver microsomes and pooled female rat (Sprague-Dawley) liver microsomes were obtained from Sigma-Aldrich (St. Louis, MO, USA). Pooled male human liver microsomes (Lot No. 2110108) and pooled female human liver microsomes (Lot No. 1210079) were obtained from XenoTech (Lenexa, Kansas City, KS, USA). 2-Methylmethcathinone (2-MMC) hydrochloride, 3-methylmethcathinone (3-MMC) hydrochloride, and 4-methylmethcathinone (4-MMC) hydrochloride standards were provided by the Amsterdam Dutch Police. All stock solutions were prepared in DMSO and stored at -80 °C until further use.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!