Eighty micrograms of each sample were reduced with 10 mM dithiothreitol (BioUltra, Sigma) for 45 min at 56 °C, alkylated with 20 mM iodoacetamide (BioUltra, Sigma) for 30 min at room temperature in the dark, and then precipitated with acetone (HPLC Plus, Sigma) overnight at −20 °C. The dried sample was resuspended in 50 mM ammonium bicarbonate (BioUltra, Sigma) and digested overnight at 37 °C with trypsin (Sequencing Grade Modified Trypsin, Promega) in a 1:50 trypsin:sample ratio. A second digestion was performed for 3 h at 37 °C, by adding trypsin in a 1:100 trypsin:sample ratio with 80% (v/v) acetonitrile (Optima LC/MS grade, Fisher Scientific). The sample was dried on a SpeedVac (ThermoSavant Scientific) and resuspended in 5% Formic acid (Optima LC/MS grade, Fisher Scientific) to perform peptide cleanup using C18 microcolumns (OMIX C18 pipette tips, Agilent), and then dried again.
Bioultra
Manufactured by Merck Group
Sourced in United States, Germany
BioUltra is a versatile lab equipment product from Merck Group. It is designed for general laboratory use, providing reliable and consistent performance in various experimental and analytical applications. The core function of BioUltra is to assist in the precise and efficient handling of samples and materials within a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Bioxtra, by Merck Group (5 mentions)
Milli q, by Merck Group (2 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (2 mentions)
96 well clear round bottom plates, by Greiner (2 mentions)
Chelex 100 resin, by Bio-Rad (2 mentions)
Spark 10m, by Tecan (2 mentions)
Exosome precipitation kit, by System Biosciences (2 mentions)
Exo fbs, by System Biosciences (2 mentions)
Hplc grade acetonitrile, by Merck Group (2 mentions)
Titrisol, by Merck Group (1 mentions)
3 aminopropyldimethylethoxysilane, by ABCR (1 mentions)
Syringe filter, by Carl Roth (1 mentions)
Poly allylamine hydrochloride pah, by Merck Group (1 mentions)
Sodium borohydride nabh4, by Merck Group (1 mentions)
L ascorbic acid bioxtra, by Merck Group (1 mentions)
Gold 3 chloride trihydrate haucl4 3h2o, by Merck Group (1 mentions)
Hexadecyltrimethylammonium bromide ctab, by Merck Group (1 mentions)
Silver nitrate agno3, by Merck Group (1 mentions)
Phosphate buffer saline pbs, by Corning (1 mentions)
Silver paste, by Ted Pella (1 mentions)
32 protocols using bioultra
1
Protein Sample Preparation for LC-MS
2
Sterilization and Plant Culture Media
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Sodium hypochlorite solution (NaOCl) EMPLURA and ethanol gradient grade for liquid chromatography LiChrosolv ® were purchased from Merck KGaA (Darmstadt, Germany). Murashige and Skoog medium (MS), plant preservative mixture (PPM), and agar were obtained from Duchefa Biochemie (Haarlem, The Netherlands). TWEEN 20 Cell Culture Tested, 6-Benzylaminopurine hydrochloride (BA) suitable for plant cell culture, indol-3-acetic acid (IAA) BioReagent, suitable for plant cell culture, sucrose for molecular biology, indole-3-butyric acid (IBA) suitable for plant cell culture, sodium carbonate (Na 2 CO 3 ) BioXtra, sodium bicarbonate (NaHCO 3 ) BioXtra, polyvinylpyrrolidone molecular weight 40,000 (PVP40), bovine serum albumin (BSA), BioReagent, suitable for cell culture, acetic acid glacial, ACS reagent, glycine, BioUltra, for molecular biology, sodium chloride (NaCl), BioXtra, ethylenediaminetetraacetic acid (EDTA), BioUltra, Triton X, BioXtra, and β-mercaptoethanol, BioUltra, for molecular biology were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Preparation of Functionalized Colloidal Particles
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All aqueous solutions have been prepared with deionized water of Milli-Q grade and a resistivity >18.2 MΩ/cm at 25 °C. Ionic strength and pH of solutions was adjusted by means of 1 M HCl, 1 M KOH (Titrisol, Merck) solutions and KCl (Bio Ultra, Sigma-Aldrich). All solutions were degassed by applying vacuum for at least 30 min before the experiment and filtered using a syringe filter with a pore size of 0.22 µm (Carl Roth GmbH & Co KG). 3-aminopropyldimethylethoxysilane (ABCR GmbH), used for cantilever modification. Sulfate modified latex particles with average diameters of 4 µm and 0.9 µm were purchased from Molecular Probes. Carboxyl modified latex particles with an average diameter of 330 nm were purchased from Invitrogen (Thermo Fisher Scientific).
4
Gold Nanoparticle Synthesis and Characterization
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Gold(iii ) chloride trihydrate (HAuCl4·3H2O, ≥99.9%, Sigma-Aldrich), sodium borohydride (NaBH4, >98%, Sigma-Aldrich), hexadecyltrimethylammonium bromide (CTAB, ≥99%, Sigma-Aldrich), sodium citrate (tribasic dehydrate, C6H5Na3O7·2H2O, ≥99%, Sigma-Aldrich), poly(allylamine hydrochloride) (PAH, M.W. 17,500 g mol−1, Sigma-Aldrich), poly(acrylic acid) sodium salt (PAA, M.W. 15,000 g mol−1, Sigma-Aldrich), silver nitrate (AgNO3, 99.0%, Sigma-Aldrich), l -ascorbic acid (BioXtra, ≥99.0%, crystalline, Sigma-Aldrich), hydrochloric acid (HCl, certified 1.0 N, Fisher Chemical), phosphate-Buffer Saline (PBS, 1×, Corning), exosome-depleted FBS (Exo-FBS, SBI), trypan blue solution (0.4%, Invitrogen), Triton X-100 solution (BioUltra, Sigma-Aldrich), exosome precipitation kit (Exo-quick, SBI). All solutions were prepared with nanopure water.
5
Quantifying Glass Dissolution Kinetics
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The mass loss assessment was performed on all glass compositions in triplicate at timepoints of 1, 12 and 24 h. For each glass sample, 0.1 g of glass powder was placed in a pre weighed 15 mL HDPE centrifuge tube with 10 mL of TRIS-buffered saline solution (BioUltra, Sigma Aldrich, Oakville, ON, Canada), capped and placed horizontally on a shaking incubator at 37 °C and agitated at 2 Hz until the desired time point.
Following incubation, the samples were centrifuged for 15 min and 300 RCF to remove residual glass from the solution. Tubes were carefully decanted to remove the supernatant, which was capped and stored at 4 °C until the time of fluoride release analysis. Test tubes with glass residuals were dried in an oven at 70 °C until constant weight was achieved (approximately 2 weeks) to determine residual glass mass and calculate the % of weight loss.
Following incubation, the samples were centrifuged for 15 min and 300 RCF to remove residual glass from the solution. Tubes were carefully decanted to remove the supernatant, which was capped and stored at 4 °C until the time of fluoride release analysis. Test tubes with glass residuals were dried in an oven at 70 °C until constant weight was achieved (approximately 2 weeks) to determine residual glass mass and calculate the % of weight loss.
6
Biocompounds Obtained via Sigma-Aldrich
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Chemicals were obtained as grade BioUltra from Sigma-Aldrich unless otherwise stated.
7
Trace Metal Contamination Removal
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Removal of contaminating metals from glassware and solutions is a critical precaution to ensure reproducibility of experiments. All glassware was treated with 3.7% hydrochloric acid for ≥12 hours, followed in some cases by ≥12 hours with 7% nitric acid to remove trace metal contamination. All solutions, buffers, and washes were prepared using Milli-Q (Millipore Sigma) ultrapure water. Solutions were prepared using BioUltra grade (Sigma-Aldrich) reagents, when available. For yeast media, addition of all components was done without the use of metal spatulas. Media were filtered through 0.2-μm membranes. Fermentative media were SC (SC medium with 2% glucose, all amino acids, and uracil and adenine), SC lacking lysine or glutamate, or minimal medium (with 2% glucose and either no amino acids or only essential auxotrophic metabolites). Agar media were prepared using acid-washed glassware.
8
Fabrication of Responsive Nanocomposite Structures
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Lithium fluoride (LiF, Sigma-Aldrich, BioUltra, ≥99.0%), hydrochloric acid (HCl, Sigma-Aldrich, ACS reagent, 37%), Ti3AlC2 MAX powders (MAX, Tongrun Info Technology Co. Ltd, China), single-walled carbon nanotubes (SWNTs, Timesnano Co. Ltd, China), sodium dodecyl sulfate (SDS, Sigma-Aldrich, >99.9%), poly(vinyl alcohol) (PVA) (Mw 67,000, Sigma-Aldrich), ethanol (Thermo Fisher, >99.5%), and DCM (J.T. Baker; 99.9%) were used as received without further purification. Biaxial PS shrink films were purchased from Grafix. Thermally responsive shrink films (with uniaxial contraction mode) were produced by EVERGREEN SCIENTIFICS. VHBTM Tape 4910 (1 inch × 36 yards) was purchased from 3 M. Silver paste was purchased from Ted Pella Inc. Commercial standard resistors were purchased from TELESKY. Deionized (DI) water (18.2 MΩ) was obtained from a Milli-Q water purification system (Merck Millipore) and used as the water source throughout the work.
9
Reagent Preparation for HPLC Analysis
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All the reagents were purchased at the highest commercial grade possible and used without any further purification, unless otherwise noted. Acetonitrile (HPLC grade, cat# 34851, Sigma-Aldrich, USA), methanol (HPLC grade, cat# A452SK-4, Fisher Scientific, USA), N,N-dimethyl acetamide (DMA) (HPLC grade, 99.5%, cat# 22916, Alfa Aesar, USA), Dimethyl sulfoxide (DMSO) (≥99.5%, cat# D5879, Sigma-Aldrich, USA), 1,1,1,3,3,3-Hexafluoroisopropyl alcohol (99.9%, cat# 00080, Chem-Impex international, Inc., USA), Triethylamine (≥99%, cat# T0886, Sigma-Aldrich, US). UltraPure distilled water (DNAse, RNAse free, cat# 10977-015, Invitrogen, USA) and sodium hydroxide solution (BioUltra, 10 M in H2O, cat# 72068, Sigma-Aldrich, USA) were used for buffer preparation. Deionized water was used for LCMS mobile phase preparation.
10
Enzymatic Hydrolysis of β-Casein for 11-mer Peptide
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In order to evaluate whether the purified protease hydrolyzes β-casein protein to obtain 11-mer peptide, an enzymatic activity assay was performed with β-casein as substrate and purified protease from CECT 7210 supernatant as enzyme. The hydrolysis was carried out in a solution containing 15 ml of protease-purified fraction (0.04 U), 30 mL of 2% β-casein solution (BioUltra, Sigma–Aldrich) and 105 mL phosphate citrate buffer (50 mM pH 6.4). After 48 h hydrolysis at 50°C, samples were boiled for 10 min and peptides formation was analyzed by HPLC. Evaluation of 11-mer formation and β-casein degradation was conducted on a HPLC (Waters 2695) with photodiode array detector (Waters 2996) and C18 column (Sunfire Waters C18 5 mm 4.6∗150 mm). Eluents were water (MilliQ quality; A) and acetonitrile (90% v/v; B), both with TFA 0.1% (v/v). Chromatographic conditions are summarized in Table 1 .
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