The non-protein bound fraction of [18F]GSK2647544 in arterial plasma (fp) was quantified using the following methodology. An aliquot of [18F]GSK2647544 was added to approximately 1 ml of plasma collected from the subject prior to tracer administration or to 1 ml Tris buffer (pH = 7.4, Sigma). Triplicate 50 μl aliquots of spiked plasma and Tris buffer were prepared for gamma counting using an automated gamma counter (Perkin Elmer, Massachusetts, US). Subsequently, triplicate 200 μl aliquots of spiked plasma and Tris buffer were pipetted into ultrafiltration units (Amicon Ultra-0.5 ml centrifugal filters, Merck Millipore) pre-treated with a 5 % TWEEN 80 solution. Filters were centrifuged at room temperature for 15 min at 10,000 rpm. At the end of centrifugation, 50 μl plasma and Tris ultrafiltrate were counted using the automated gamma counter and fp was calculated as the ratio of ultrafiltrate to total activity concentrations corrected for non-specific filter binding. There was no evidence of [18F]GSK2647544 being metabolised during this procedure.
Tris buffer
Manufactured by Merck Group
Sourced in United States, Germany, India, United Kingdom, Italy, France
Tris buffer is a widely used chemical solution in biochemistry and molecular biology laboratories. It serves as a buffering agent, maintaining a stable pH environment for various biological and chemical reactions. The core function of Tris buffer is to maintain a consistent and controlled pH level within samples, enabling researchers to optimize the performance of enzymes, proteins, and other biomolecules during experiments and analysis.
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Lab products found in correlation
Sodium chloride (nacl), by Merck Group (18 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (14 mentions)
Bovine serum albumin (bsa), by Merck Group (12 mentions)
Acetic acid, by Merck Group (12 mentions)
Triton x 100, by Merck Group (8 mentions)
Hydrochloric acid (hcl), by Merck Group (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (6 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (6 mentions)
Sodium hydroxide (naoh), by Merck Group (6 mentions)
Tween 20, by Merck Group (6 mentions)
Trichloroacetic acid, by Merck Group (6 mentions)
Cacl2, by Merck Group (5 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (5 mentions)
Phosphate buffered saline (pbs), by Merck Group (5 mentions)
Sulforhodamine b, by Merck Group (5 mentions)
Phosphate buffer, by Merck Group (5 mentions)
Sucrose, by Merck Group (5 mentions)
Phosphate buffer saline (pbs), by Merck Group (5 mentions)
Penicillin, by Merck Group (5 mentions)
148 protocols using tris buffer
1
Quantifying Free Fraction of [18F]GSK2647544
2
Intracellular ALP Activity Measurement
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The activity of the intracellularly generated ALP was analyzed after 1, 3, 7, 14, 21, and 28 days of stimulation, each 20 h after the last stimulation interval started. The cells were washed twice using TRIS buffer (50 mM, pH = 8.0) lysed with 1% Triton X and 1% phenylmethylsulfonyl fluoride (both: Merck, Darmstadt, Germany) for 10 min. A solution containing 10 mM 4-Nitrophenylphosphat (AppliChem, Darmstadt, Germany), 100 mM 2-amino-2-methyl-1,3-propanediol (Sigma-Aldrich, Munich, Germany), and 5 mM magnesium chloride (Merck, Darmstadt, Germany) was added to the lysate and incubated over 1 h at 37°C and 5% CO2. The reaction was stopped with a 2 M sodium hydroxide solution, and the absorption was measured at 405 nm with multimode plate reader Infinite 200 pro (Tecan Group Ltd., Maennedorf, Switzerland). A blank served as an internal control and was subtracted from each value.
3
Protocols for Neuronal Cell Culture and Analysis
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Materials were acquired from the following sources: Gold chloride, poly (ethylene glycol) (PEG), poly-D-lysine (PDL), bovine serum albumin (BSA), colchicine and paraformaldehyde (PFA): Sigma-Aldrich, USA; Trisodium citrate dihydrate, papain, nitric acid, hydrogen peroxide, hydrochloric acid, TRIS-buffer, triton X-100, di-sodium hydrogen phosphate and potassium dihydrogen phosphate: Merck Chemicals, USA; Collagenase-II, dispase-II, fetal bovine serum (FBS), penicillin–streptomycin mix and trypsin–EDTA: Gibco, USA; Hank's buffered saline solution (HBSS), Ham’s F-12 medium and Dulbecco’s modified Eagle’s medium (DMEM): Lonza Chemicals, USA; Isolectin B4, DyLight 594-IB4, FITC-IB4, DyLight 488 horse anti-mouse IgG antibody and Vectashield hardset mounting medium: Vector Laboratories, USA; Glutaraldehyde (EM grade), osmium tetroxide (OsO4), sodium cacodylate trihydrate and EMBed-812 embedding kit: Electron Microscopy Sciences, USA; Hoechst 33342: Santa Cruz Biotechnology, Inc. USA; Anti β-III tubulin monoclonal antibody: Cell Signaling Technology, Inc USA; Anti PGP9.5 antibody: Abcam Plc UK; Microfluidic devices (SND 150): Xona Microfluidics, LLC, USA.
4
Synthesis and Characterization of SBF
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The SBF solution was prepared according to Kokubo's specifications.32 Some reagents such as NaCl, NaHCO3, KCl, K2HPO4·3H2O, MgCl2·6H2O, CaCl2, trishydroxymethyl aminomethane [Tris buffer, (CH2OH)3CNH2], and 1 N HCl, were purchased from Merck Inc. The SBF solution was prepared by dissolving NaCl, KCl, NaHCO3, MgCl2·6H2O, CaCl2 and KH2PO4 (analytical grade) in distilled water and buffered at pH = 7.25 with Tris buffer and HCl 1 N at 37 °C.33
5
Phytochemical Extraction and Bioactivity
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The solvents used for extraction of plant material were of HPLC grade. Tris buffer were purchased from Merck (Darmstadt, Germany), porcine pancreatic lipase, ascorbic acid, starch, acarbose, DPPH (2,2-diphenyl-1-picrylhydrazyl), PNPP (p-nitrophenyl palmitate) and orlistat were bought from Sigma-Aldrich (USA) and porcine pancreatic α-amylase from MP Biochemicals (Illkirch, France). All the chemicals used in the experiments were of analytical grade.
6
Polymeric Nanocarrier Formulation and Evaluation
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PLGA Resomer® RG503H (50:50; Mw 24,000–38,000), ethyl acetate, Pluronic®F127, coumarin-6 (C6) (Mw 350.43), phosphate buffered saline (PBS), acetic acid, sulforhodamine B (SRB), trypan blue, ribonuclease A (RNase) from bovine pancreas (Mw 13,700; solution of 50% glycerol), propidium iodide (Mw 668.39, purity ≥ 94%) and Triton XTM-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cholecalciferol (vitamin D3, Mw 384.65, purity ≥ 99%) was purchased from Alfa Aesar (Karlsruhe, Germany). Calcitriol (Rocaltrol, Mw 416.64, purity ≥ 99%) was purchased from Selleck Chemicals (Munich, Germany). Uranyl acetate (dehydrate, 424.146 g/mol) was purchased from Electron Microscopy Sciences (Hatfield, UK). Dulbecco's Modified Eagle medium (DMEM) and Roswell Park Memorial Institute medium (RPMI) were acquired from Invitrogen Co. (Scotland, UK). Trichloroacetic acid (TCA) and Tris buffer were acquired from Merck (Darmstadt, Germany). SlowFade® Gold Antifade Mountant with DAPI and LysoTracker® Deep Red were purchased from Molecular Probes (Invitrogen Co., Scotland).
7
Synthesis and Characterization of Protein Conjugates
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HSA (≥96%, free fatty acid), β-CyD, β-D(+) glucose, sodium Alg and sodium bicinchoninate (BCA) were purchased from Sigma (USA). Membrane filters (0.2 μm pore size, 25 mm in diameter) and dialysis tubings (cut off 10,000 MW) were from Whatman (UK). Tre, DTT, Tris buffer and sodium azide were from Merck (Germany). All other materials were of analytical grade. All solutions were prepared with deionized water.
8
Cultivating Stratified Cells for Vascular Research
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One or two SGE were placed onto the abluminal side of the adventitia and the luminal side of the intima and media. The three different dEAC layers with SGE were cultivated for 5 days in a medium consisting of Panserin 401 (PAN-Biotech, Aidenbach, Germany) supplemented with insulin (8.7 µg/mL; Biochrom, Berlin, Germany), penicillin (30 U/mL; Biochrom), glucose (0.15%; B. Braun Melsungen, Melsungen, Germany), phosphate-buffered saline (PBS; Ca2+/Mg2+-free PBS; 0.172 mg/mL; Gibco by Life Technologies, Carlsbad, CA, USA), HEPES buffer solution (23.43 µM; Invitrogen, Carlsbad, CA, USA) and N-2 supplement (0.1 µL/mL; Invitrogen) in an incubator (CB 150 E3; Binder, Tuttlingen, Germany) at 37°C, 5% CO2 and humidity of 95%. The serum-free medium was conditioned with 10% fetal calf serum (FCS). At the end of the experiments, dEAC layers with SGE were fixed with a 4% paraformaldehyde (PFA) solution per well for 1 h at 4°C and washed two times using hydroxymethyl-aminomethane (TRIS) buffer (Merck) and stored at 4°C.22 (link) The experiments were repeated in triplicate with SGE from different animals for each of the layers in one-well plate.
9
Tyrosinase Modulation by Santalol
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The target enzyme, tyrosinase was obtained from an edible mushroom (Agaricus bisporus). The test compounds, santalol (CAS no. 77-42-9, 93% pure), dimethyl-sulfoxide (DMSO), and 3, 4-Dihydroxyphenylalanine (L-DOPA), were bought from Sigma-Aldrich. All other reagents such as phosphate buffer, Tris buffer, sodium chloride, SDS, etc. of analytical grades were purchased from Merck ltd (Bengaluru, India).
10
Acinetobacter baumannii DNA Hybridization Protocol
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All solutions were prepared with doubly distilled deionized water purchased from Shahid Ghazi Pharmaceutical Company (Tabriz, Iran). DNA oligonucleotide sequences were acquired from Takapouzist Co. (Iran) Tris buffer, NaCl and Sodium acetate were obtained from Merck (Darmstadt, Germany). Ethyl acetate obtained from Sharlo Company (Spanish). Source of Acinetobacter baumannii is OMPA gene (IX87-RS045). Acinetobacter baumannii complete oligonucleotide sequences (5ʹ SH-TTG TGA ACT ATT TAC GTC AGC ATG C 3ʹ), GC ratio: 40%, basecount: 25, Tm: 59.4. Complementary target sequence (5ʹ GCA TGC TGA CGT AAA TAG TTC ACA A 3ʹ), basecount: 25,Tm: 59.4, GC ratio: 40%. Single nucleotide mismatch target sequence (5ʹ GTA TGC TGA CGT AAA TAG TTC ACA A3ʹ), GC ratio: 36%, Tm: 56.7, basecount: 25. Two nucleotide mismatch target sequence (5ʹ GTA TGA TGA CGT AAA TAG TTC ACA A 3ʹ), basecount: 25, GC ratio: 32%, Tm: 54.6 [16 ]. The oligonucleotide stock solutions were diluted with 0.1 M Tris-HCl buffer, pH 7.4 solution (Tris). Dithiothreitol (DTT) (Sigma-Aldrich company united-states) solved in Tris-HCl and employed as a redox indicator for revealing DNA hybridization. All the above solutions were kept at 4 °C before use. DTT solution containing 10 mM Sodium acetate and 500 mM DTT, pH (5.2) was prepared and kept at 4 °C.
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