Anhydrous 1,6-hexamethylene bis(chloroformate) (95%) and trimethylolpropane tri(chloroformate) (97%) were purchased from Manchester Organics Ltd (UK) and used as received. 4-Dimethylaminopyridine (DMAP), anhydrous pyridine, anhydrous dimethylsulfoxide (DMSO) and anhydrous dichloromethane (DCM) were purchased from Merck (UK) and used as received. N,N′-Dimethylformamide (DMF), ethanol, ethyl acetate, hexane, CDCl3, and magnesium sulfate (MgSO4) were purchased from Fischer Scientific, whilst anhydrous ethanol and hexyl chloroformate was purchased from Sigma Aldrich. Emtricitabine (FTC) was purchased from WISCHEM (China) and used without further purification. Male and female human plasma and the dipotassium salt of ethylenediaminetetraacetic acid (K2-EDTA) were purchased from BioIVT and the plasma samples were pooled. Benzil was purchased from Merck (Germany). Pooled, mixed gender, adult human liver microsome and phosphate buffered saline (PBS) at pH 7.4 was purchased from ThermoFisher Scientific (USA). Carboxylesterase 1 (CES1) specific activity assay kit was purchased from Abcam (UK).
Anhydrous dimethyl sulfoxide
Manufactured by Merck Group
Sourced in United States, Germany, Italy, United Kingdom, France, Austria
Anhydrous dimethyl sulfoxide (DMSO) is a clear, colorless, and aprotic solvent. It has a high boiling point and is miscible with water and a wide range of organic solvents. DMSO is commonly used as a solvent and reaction medium in various laboratory and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Triethylamine, by Merck Group (13 mentions)
Dextran, by Merck Group (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Anhydrous chlorobenzene, by Merck Group (6 mentions)
Dichloromethane, by Merck Group (6 mentions)
2 methoxypropene, by Merck Group (5 mentions)
4 dimethylaminopyridine, by Merck Group (5 mentions)
Methanol, by Merck Group (5 mentions)
Phosphate buffered saline (pbs), by Merck Group (5 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions)
Methacrylic anhydride, by Merck Group (4 mentions)
Pyridinium p toluenesulfonate, by Merck Group (4 mentions)
Anhydrous acetonitrile, by Merck Group (4 mentions)
Sodium acetate, by Merck Group (4 mentions)
Anhydrous n n dimethylformamide, by Merck Group (4 mentions)
Hydrochloric acid (hcl), by Merck Group (4 mentions)
Ethanol, by Merck Group (4 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (4 mentions)
Anhydrous dichloromethane, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Corning (3 mentions)
93 protocols using anhydrous dimethyl sulfoxide
1
Synthesis and characterization of FTC prodrugs
2
Preparation of Amyloid-Beta Oligomers
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Aβ oligomers were prepared from Aβ1–42 peptide, as described by Klein [24 (link)]. Briefly, ice-cold Aβ1–42 peptide was dissolved in an ice-cold solution of hexafluoroisopropanol (HFIP; Sigma-Aldrich, USA) to enable dissociation of any Aβ aggregates and a final concentration of 1 mM HFIP was made. The Aβ–HFIP mixture was kept aside for 1 h incubation period. Then, the solution was kept on ice and vortexed and aliquoted into Eppendorf tubes, and HFIP was allowed to evaporate by leaving the tubes overnight. The lyophilized sample was again suspended in anhydrous dimethyl sulfoxide (Merck, USA) to make a 5 mM stock solution of Aβ oligomer. The resulting mixture containing insoluble fibrillar aggregates and soluble oligomeric Aβ peptides was processed with Ham’s F-12 medium. Finally, the supernatant solution containing oligomeric forms of Aβ peptides was separated and stored at 4℃ until required.
3
Chondroitin Sulfate Hydrogel Synthesis
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All used reagents were of analytical grade. Type A gelatin (Gel) from porcine skin (~300 bloom), chondroitin sulfate A sodium salt, anhydrous dimethyl sulfoxide (DMSO), deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated water (D 2 O), methacrylic anhydride (MAA), nicotinamide (Nic), ammonium peroxydisulfate (APS), N,N,N ′ ,N ′ -tetramethylethylenediamine (TEMED), glycidyl methacrylate (GMA), 4-dimethyl aminopyridine (DMAP), tetrabutylammonium bromide (TBAB), sodium chloride, dialysis membranes (cut-off 12-14 kDa) and L-glutamine were purchased from Merck, (Milan, Italy). Spongostan TM Dental was purchased from Ethicon (Somerville, NJ, USA). Absolute ethanol (EtOH), 37% w/w hydrochloric acid (HCl), monobasic potassium phosphate (KH 2 PO 4 ) and sodium hydroxide in pellets (NaOH) were purchased from Carlo Erba (Milan, Italy). Vybrant ® CDFA SE cell tracer kit, alamarBlue™ cell viability reagent and Primocin TM were purchased from Invitrogen (ThermoFisher Scientific, Waltham, MA, USA) Fetal Bovine Serum (FBS) from Gibco (Buffalo, NY, USA), Dulbecco's Phosphate Buffered Saline (PBS, 1X) from Aurogene (Rome, Italy), DMEM, alpha MEM culture medium and penicillin-streptomycin solution (10,000 U/mL) from Euroclone (Milan, Italy).
4
Fluorescent Labeling of Therapeutic Antibodies
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The antibodies Infliximab (Remicade®), adalimumab (Humira®), vedolizumab (Entyvio®) and ustekinumab (Stelara®) were investigated. All antibodies were acquired commercially. Stock solutions were made by dissolving lyophilized antibody in water for injections (B. Braun, Melsungen, Germany), or extracting solution from syringes or pens into Eppendorf cups, as appropriate. All stock solutions were stored refrigerated at 2–8 °C during conjugation experiments. Further details on the antibodies are displayed in Table S1 .
IRDye 800CW, IRDye 680LT (both LI-COR Bioscience, Lincoln, NE, USA) and ZW800-1 (Curadel, Marlborough, MA, USA) were selected based on the presence of a peak in their fluorescence spectrum in the 690 nm to 800 nm range and availability as NHS-ester. Chemical details on the used dyes are displayed inTable S2 . All dyes were dissolved in anhydrous dimethyl sulfoxide (Merck, Darmstadt, Germany) and stored at −80 °C until use.
IRDye 800CW, IRDye 680LT (both LI-COR Bioscience, Lincoln, NE, USA) and ZW800-1 (Curadel, Marlborough, MA, USA) were selected based on the presence of a peak in their fluorescence spectrum in the 690 nm to 800 nm range and availability as NHS-ester. Chemical details on the used dyes are displayed in
5
Erythrocyte Calcium Dynamics Quantification
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Intraerythrocytic Ca2+ concentrations were assessed by FLUO4 AM staining. This Ca2+-sensitive dye was purchased from Becton Dickinson (USA). Its 5 mM stock solution was in anhydrous dimethyl sulfoxide (Sigma Aldrich, USA) that was prepared and used to stain 2 μl of erythrocytes dissolved in 98 μl PBS to obtain 2.5 μM dye working solutions. The same volumes (400 μl) of PBS were added after the incubation lasted for 30 min in the dark. The FLUO4 fluorescence was acquired at 520 nm after excitation with the 488 nm laser.
Fluorescence of Annexin V-FITC, DCF, and FLUO4 was expressed in arbitrary units (a.u.). Erythrocyte suspensions without the corresponding dyes were considered negative controls. Erythrocytes incubated with H2O2 (0.1 mM) were used as positive controls.
Fluorescence of Annexin V-FITC, DCF, and FLUO4 was expressed in arbitrary units (a.u.). Erythrocyte suspensions without the corresponding dyes were considered negative controls. Erythrocytes incubated with H2O2 (0.1 mM) were used as positive controls.
6
Synthetic Polymeric Biomaterials Fabrication
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Sebacic acid (99.0%), malonic acid (99.0%), glutaric acid (99.0%), polycaprolactone triol (PCL triol, 300 Da), glycerol (99.0%), PEG (200, 1000, 2000 Da), N,N′-diisopropylcarbodiimide (DIC, 99.0%), 4-dimethylaminopyridine (DMAP, 99.0%), anhydrous N,N-dimethylformamide (DMF, 99.8%), anhydrous dimethyl sulfoxide (DMSO, 99.9%), anhydrous dichloromethane (DCM, 99.8%), deuterochloroform (99.96 atom% D), sodium octyl sulfate (SOS, 95.0%), dexamethasone (98.0%), fluorescein isothiocyanate isomer I (FITC, 90.0%), fluorescein sodium salt, hexamethylene diisocyanate (HMDI, 99.0%), dibutyltin dilaurate (Sn(II), 95.0%), and PBS (pH 7.4) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyanine5.5 carboxylic acid (Cy5.5, 95.0%) was purchased from Lumiprobe Corporation (Hallandale Beach, FL, USA). Tetrodotoxin (TTX) was obtained from Abcam (Cambridge, MA, USA). TTX ELISA kits were purchased from Reagen LLC (Moorestown, NJ, USA). Dulbecco’s minimum essential medium (DMEM), fetal bovine serum (FBS), and Penicillin Streptomycin was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
7
Equine Skeletal Muscle Mb Crosslinking
8
Gelatin-based Biomaterial Synthesis
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Example 1
Gelatin (type A from porcine skin, >300 bloom), 2-iminothiolane hydrochloride (TR), calcium peroxide (CaO2), anhydrous Dimethyl Sulfoxide (anhydrous DMSO) and collagenase type II were purchased from Sigma Aldrich (St. Louis, Mo., USA).
Dulbecco's Modified Eagle's Medium; DMEM), penicillin-streptomycin (P/S), trypsin/EDTA and DPBS (Dulbecco's phosphate buffered saline; DPBS) solution were purchased form Gibco (Grand Island, N.Y., USA), Fetal Bovine Serum (FBS) was purchased from Research and Diagnostic Technology. Further, EGM-2 Single quots medium (Endothelial Cell Growth Medium; EGM) was supplied from Lonza. Moreover, Cell Proliferation reagent WST-1 was purchased from Roche Diagnostics, and Live/Dead Viability/Cytotoxicity Kit was purchased from Life science.
Other chemicals and solvents were used without additional purification.
9
Preparation of Aβ Peptide Monomers and Oligomers
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The Aβ1-42 (GenicBio Limited) and Aβ42-1 peptide (AnaSpec) peptides were dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP; Sigma-Aldrich) to a final concentration of 5 mM and placed under a chemical hood overnight. The next day, HFIP was further evaporated using a SpeedVac Concentrator for 1 hour. Monomer Aβ (5 mM) was prepared by dissolving Aβ peptide in anhydrous dimethyl sulfoxide (Sigma-Aldrich). The oligomeric Aβ and control peptides were prepared by diluting the monomer Aβ solution in Dulbecco’s modified Eagle’s medium (DMEM)/F12 and then incubating at 4°C for 24 hours.
10
Biomimetic Hydrogel for Cell Culture
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