The PDA solution was prepared by dissolving 1 mg of L-DOPA in 1 mL of 10 mM Tris buffer base (pH 8.5; Sigma-Aldrich, St. Louis, MI, USA). Then, GO dispersion was added to the PDA solution (1 mg/mL) under magnetic stirring at room temperature for 24 h. The final concentration of GO in the PDA solution was 0.1, 0.5, or 1 mg/mL for individual experiments. The polystyrene (PT) culture surface was coated with PDA/GO composite solution overnight at room temperature and washed three times with sterile phosphate-buffered saline (PBS). Then, the PDA/GO-modified surfaces were dried in a vacuum oven.
Tris base buffer
Manufactured by Merck Group
Sourced in United States
Tris base buffer is a common laboratory buffer solution used to maintain a specific pH range in various biochemical and molecular biology applications. It is composed of the organic compound tris(hydroxymethyl)aminomethane (Tris) and serves as a pH stabilizer. The core function of Tris base buffer is to provide a stable pH environment for the effective operation of enzymes, proteins, and other biomolecules during experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Acetic acid, by Merck Group (2 mentions)
Proteomics grade trypsin, by Merck Group (1 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Ammonium bicarbonate, by Merck Group (1 mentions)
Iodoacetamide, by Merck Group (1 mentions)
Ethylenediaminetetraacetic acid, by Merck Group (1 mentions)
Tris 2 carboxyethyl phosphine, by Merck Group (1 mentions)
Hyclone molecular grade water, by GE Healthcare (1 mentions)
Trichloroacetic acid tca, by Merck Group (1 mentions)
Sulforhodamine b srb, by Merck Group (1 mentions)
Emax plus, by Molecular Devices (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Synergy 2, by Agilent Technologies (1 mentions)
5 protocols using tris base buffer
1
Polydopamine-Graphene Oxide Composite Coating
2
Fucoidan-Enhanced L-DOPA Coating for Cell Culture
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The PDA solution was prepared by dissolving 1 mg of L-DOPA in 1 mL of 10 mM Tris buffer base (pH 8.5; Sigma-Aldrich). Fucoidan was added to the PDA solution (1 mg/mL), and the mixture was stirred at room temperature for 24 h. The final concentration of fucoidan in the PDA solution was 0.5 or 1 µg/mL for each experiment. The PT culture surface was layered with a fucoidan/PDA composite solution overnight at room temperature and washed three times with sterile phosphate-buffered saline (PBS). Fucoidan/PDA-coated surfaces were then dried in a vacuum oven.
3
Proteomics Sample Preparation Protocol
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Hyclone molecular grade water was obtained from GE health care. Sodium chloride, formic acid, acetonitrile, and methanol were obtained from Thermo Fisher Scientific. Tris base buffer, ethylenediaminetetraacetic acid, acetic acid, ammonium bicarbonate, tris(2-carboxyethyl) phosphine, iodoacetamide, and proteomics grade trypsin were obtained from Sigma-Aldrich.
4
Sulforhodamine B Assay for Cell Viability
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Cell viability was assessed with a sulforhodamine B (SRB; Sigma) assay. SRB can bind to basic amino acids in proteins. The amount of dye represents the number of cells. Briefly, cells ( ) were seeded in a 96-well culture plate and incubated overnight at 37°C. HK-2 cells were treated with PS-MPs (Life Technologies) at 0, 0.05, 0.1, 0.2, 0.4, or for 1, 2, or 3 d. The cells were fixed with ice-cold 10% trichloroacetic acid (TCA; Sigma) at 4°C for at least 1 h or overnight. The TCA was removed, and the cells were washed twice with distilled water. Then, 0.1% SRB in 1% acetic acid (Sigma) was added, and the cell suspension was incubated at 25°C for 1 h. The cells were washed twice with 1% acetic acid and dried at 60°C in an oven for 30 min. Finally, the adhered cells were dissolved in Tris base buffer (Sigma), and the plate was shaken for 30 min. The absorbance of the dye suspension was measured at a wavelength of in an enzyme-linked immunosorbent assay (ELISA) reader (EMax Plus; Molecular Devices).
5
Evaluating Paclitaxel Cytotoxicity with Verapamil
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To determine cell viability to PTX after Verapamil incubation, a Sulforhodamine B (SRB) assay was performed. Briefly, 5 × 106 cells/well were seeded in 96-well plates and incubated at 37°C and 5% CO2. After 24 h, cells were treated with 10-fold serial dilutions of PTX, in the presence or absence of 10 µM Verapamil and incubated under the same conditions. In parallel, cells were treated with equivalent amount of vehicle (DMSO) up to 0.25% concentration. Then, cells were fixed with 50% (m/v) trichloroacetic acid (Merck Millipore, Darmstadt, Germany) for 1 h, washed with distilled water, and stained with 0.1% (m/v) SRB in acetic acid (Sigma-Aldrich) for 30 min at RT. After washing with 1% (v/v) acetic acid aquae solution (Merck Millipore), plates were left to dry at RT followed by SRB complex solubilization with 10 mM Tris-Base buffer (Sigma-Aldrich) for 30 min. Absorbance was measured (515 nm wavelength) using a Bio Tek SynergyTM 2 multi-mode microplate reader. The IC50 of PTX, in the presence or absence of Verapamil, was determined as described above in Section 2.3.
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