Tris hcl

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Canada

Tris-HCl is a chemical buffer solution commonly used in biotechnology and molecular biology applications. It is a mixture of tris(hydroxymethyl)aminomethane (Tris) and hydrochloric acid (HCl), which is used to maintain a specific pH range in various laboratory procedures.

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Lab products found in correlation

104 protocols using tris hcl

Oligonucleotide Hybridization Protocol

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The TRIS-HCl, HCl, and NaOH were obtained from Fisher Scientific (Waltham, MA, USA); EDTA and NaCl were obtained from Sigma-Aldrich (St. Louis, MO, USA). Oligonucleotides and nuclease free water were obtained from Integrated DNA Technologies (IDT, Coralville, IA, USA). Oligonucleotide sequences are included in Table 1. Heat Inactivated bovine serum was obtained from Life Technologies (Staten Island, NY, USA).
Hybridization buffer (10 mM TRIS-HCl, 1 mM EDTA, 50 mM NaCl, pH 8.0) was prepared in our laboratory.

Adegbenro A., Coleman S, & Nesterova I.V. (2021). Stoichiometric approach to quantitative analysis of biomolecules: the case of nucleic acids. Analytical and bioanalytical chemistry, 414(4), 1587-1594.

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ATP-Binding Aptamer Characterization Protocol

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Jurkat, Clone E6.1 (T lymphocyte), cells were purchased from the American Type Culture Collection (ATCC). The cell line was cultured in HyClone RPMI-1640 (+25mM HEPES, +L-Glutamine) medium supplemented with 100 units/mL penicillinstreptomycin 1% (Corning), 1% MEM Non-Essential Amino Acids (Gibco) and 10% fetal bovine serum (Heat Inactivated, Gibco). All cell lines were routinely evaluated on a Flow Cytometer (FACScan, Becton Dickinson) for the expression of CD marker using anti-hCD3ε (PE-conjugated Mouse IgG1, R&D Systems) antibody to authenticate the cell line. All conformational assays were tested using Adenosine-5'triphosphate, ATP, from a stock solution of 100 mM (ThermoFisher). The specificity assay was performed using 2 mM of Adenosine-5'-triphosphate, ATP, Uridine-5'triphosphate, UTP, Cytidine-5'-triphosphate, CTP, and Guanosine-5'-triphosphate, GTP, from a stock solution of 100mM (ThermoFisher). All aptamer solutions were prepared in 10 mM Tris-HCl (Thermo Scientific) adjusted to pH=8.4, with 6 mM MgCl2 (Sigma Aldrich) from a stock solution of 1M Tris-HCl (ThermoFisher). All DNA sequences were ordered HPLC-purified from Integrated DNA Technologies (IDT) and dual-modified with 6-Carboxyfluorescein (6-FAM) and Iowa Black Fluorescence Quencher (IBFQ) at the 5' and 3', respectively.

Boykoff N., Mallikaratchy P., Lenn J., & Freage L. (2021). A bispecific aptamer sensor towards T-cell leukemia detection in the tumor microenvironment.

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TRIM25-ZAP Interaction Analysis in IAV Infection

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HEK293 TRIM25 KO cells transfected with plasmids expressing T7-tagged TRIM25 and untagged ZAP were infected with the IAV PR8 R38K41A strain at an MOI of 5 for 6 h. The cells were disrupted by sonication and the extracts pre-cleared by incubation with 20 μl protein A agarose for 30 min. Anti-T7 antibody (69048 Merck Millipore) was coupled to 30 μl protein A dynabeads (Thermo Fisher Scientific) and incubated with the extracts for 1 h. The bound proteins were separated on a 4–12% SDS polyacrylamide gel and analyzed by western blotting. Washes of beads were done in Buffer G (20 mM Tris–HCl pH 7.5, 137 mM NaCl (Fisher Scientific), 1 mM EDTA, 1% Triton X-100 (Fisher Scientific), 10% glycerol, 1.5 mM MgCl₂ (Fisher Scientific), 1 mM DTT, 0.2 mM PMSF (Thermo Fisher Scientific).

Choudhury N.R., Trus I., Heikel G., Wolczyk M., Szymanski J., Bolembach A., Dos Santos Pinto R.M., Smith N., Trubitsyna M., Gaunt E., Digard P, & Michlewski G. (2022). TRIM25 inhibits influenza A virus infection, destabilizes viral mRNA, but is redundant for activating the RIG-I pathway. Nucleic Acids Research, 50(12), 7097-7114.

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Mica Cleaving for Collagen Fibrils

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Freshly and uniformly
cleaving mica is a required step in assembling reproducible collagen
fibril fields. A 15 mm × 15 mm piece of muscovite mica (highest
grade VI, Ted Pella, Redding, CA, USA) was freshly cleaved using tape.
A flat layer of the mica that remained on the tape was discarded.
Collagen type I was diluted in the buffer solution to the proper concentration
and the buffer solution was incubated on the mica at room temperature.
After incubation the collagen solution was washed with distilled water,
the mica was laid against the edge of a tissue culture dish, and the
mica was allowed to dry overnight and used the next day. There were
four different buffer solutions. Buffer solution 1 consisted of 50
mM Tris-HCl (Fisher Scientific, Hampton, New Hampshire, USA) and 200
mM KCl (Fisher Scientific) at pH 9.2. Buffer solution 2 consisted
of 50 mM Tris-HCl and 200 mM KCl at pH 7.5. Buffer solution 3 consisted
of 50 mM citric acid-sodium citrate (Fisher Scientific) and 200 mM
KCl at pH 4.3. Buffer solution 4 consisted of 50 mM citric acid-sodium
citrate and 0 mM KCl at pH 4.2.

Wang J., Petefish J.W., Hillier A.C, & Schneider I.C. (2014). Epitaxially Grown Collagen Fibrils Reveal Diversity in Contact Guidance Behavior among Cancer Cells. Langmuir, 31(1), 307-314.

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Measuring sGC Activity via cGMP Assay

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30 μg of PA protein was measured using the Bradford method. Samples were incubated in a reaction mixture of 50 mM Tris-HCl (pH 7.5, Fisher Scientific), 4 mM MgCl₂ (Fisher Scientific), 0.5 mM 3-isobutyl-1-methylxanthine (Enzo), 7.5 mM creatine phosphate (Sigma-Aldrich), 0.2 mg/mL creatine phosphokinase (Sigma-Aldrich), 1 mM sodium nitroprusside (Sigma-Aldrich), and 1 mM GTP (Sigma-Aldrich) at 37°C. After 10 minutes, the reaction was terminated using HCl (Sigma-Aldrich) to a final concentration of 0.1 N. Samples were dried using a Speed-Vac, and pellets were resuspended in 100–200 μL of cGMP EIA buffer (Cayman Chemical, Ann Arbor, MI). cGMP levels in the samples were measured in duplicate using a commercially available EIA kit (Cayman Chemical). Results were measured using a Bio-Rad iMark automated plate reader (Hercules, CA) at 405 nm [6 (link), 23 (link)]. sGC activity results are shown as pmol cGMP/minute/mg total protein.

Perez M., Lee K.J., Cardona H.J., Taylor J.M., Robbins M.E., Waypa G.B., Berkelhamer S.K, & Farrow K.N. (2017). Aberrant cGMP signaling persists during recovery in mice with oxygen-induced pulmonary hypertension. PLoS ONE, 12(8), e0180957.

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Synthesis and Characterization of C6T iM Oligonucleotide

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The
iM-forming oligonucleotide (C6T; Figure 2) used was synthesized
by and purchased from Midland Certified Reagent Co., Inc. (Midland,
TX). C6T is a good model iM because it undergoes a simple two-state
transition from folded to unfolded form.^{36 (link)} C6T iM oligonucleotide stock was stored in a 10 mM Tris, 1 mM ethylenediaminetetraacetic
acid (EDTA) buffer at pH 8.0 in a −20 °C freezer. The
sodium cacodylate, Tris-HCl, and EDTA used to create buffer solutions
were purchased from Fisher Scientific (Pittsburgh, PA). Putrescine,
spermidine, and spermine were purchased from Sigma Aldrich (St. Louis,
MO).

Molnar M.M., Liddell S.C, & Wadkins R.M. (2019). Effects of Polyamine Binding on the Stability of DNA i-Motif Structures. ACS Omega, 4(5), 8967-8973.

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Epitaxial Growth of Collagen Fibrils on Mica

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Collagen fibrils were epitaxially grown on 15 mm × 15 mm pieces of muscovite mica (highest grade VI, Ted Pella, Redding, CA, USA) that were freshly cleaved using tape.^{52 (link)} Rat tail collagen type I was diluted (BD Bioscience, 10 μg mL⁻¹) in the buffer solution consisted of 50 mM Tris–HCl (Fisher Scientific) and 200 mM KCl (Fisher Scientific) at pH 9.2. After incubation of 18 h the collagen solution was washed with deionized water, the mica was laid against the edge of a tissue culture dish and the mica was allowed to dry overnight and was used the next day.

Wang J., Boddupalli A., Koelbl J., Nam D.H., Ge X., Bratlie K.M, & Schneider I.C. (2018). Degradation and Remodeling of Epitaxially Grown Collagen Fibrils. Cellular and Molecular Bioengineering, 12(1), 69-84.

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Fluorescent Labeling of miRNAs

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miRNAs, DNA probes labeled with AlexaFluor 594, and IDTE buffer (10 mM Tris, 0.1 mM ethylenediaminetetraacetic acid, pH 7.5) were purchased from Integrated DNA Technologies (Coralville, IA). miRNAs and probes (Table S1) were reconstituted in 1x IDTE and stored at −20 °C. Pluronic F-127 (PF-127), magnesium chloride, ammonium acetate, and rhodamine 6G were obtained from Millipore Sigma (Burlington, MA). Tris-HCl and glycine were purchased from Fisher Scientific (Pittsburgh, PA). All solutions were prepared with 18.2 MΩ·cm ultrapure water from an ELGA LabWater Purelab Classic (High Wycombe, UK).

Reddy S.T., Stoker A.W, & Bissell M.J. (1991). Expression of Rous sarcoma virus-derived retroviral vectors in the avian blastoderm: potential as stable genetic markers.. Proceedings of the National Academy of Sciences of the United States of America, 88(23), 10505-10509.

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Covalent Peptide Labeling with Fluorescent Dye

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In brief: Dye was covalently attached by the Sulphur of the Cys in the peptide with Tris buffer solution at room temperature.
BDP 630/650 maleimide was purchased from Lumiprobe (Hunt Valley, MD, USA); Tris HCl was purchased from Fisher Bioreagents (Shanghai, China); Tris base was purchased from Fisher Bioreagents (Pittsburgh, PA, USA). The peptide CVD-205 (CH₃-C(O)-CGLQRMVLVDLKGGYGRKKRRQRRR-NH₂) (Figure S3) was dissolved in Tris buffer (100 mM (pH 7.4)) to 4 mM. The dye was dissolved in ACN to 3 mM. The dye solution was added to the peptide solution in equal volumes with proper shaking at 700 rounds per minute (rpm) overnight at room temperature.

Ben-Uliel S.F., Zoabi F.H., Slavin M., Sibony-Benyamini H., Kalisman N, & Qvit N. (2022). De Novo Development of Mitochondria-Targeted Molecular Probes Targeting Pink1. International Journal of Molecular Sciences, 23(11), 6076.

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Profiling Serine Hydrolase Activities

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TAF, SBV, RDV, and telaprevir were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Recombinant human CatA (rhCatA), CathepsinL (rhCatL), and CES1 (rhCES1) were purchased from R&D Systems (Minneapolis, MN, USA). Recombinant human lysophospholipase 1 (rhLYPLA1) was purchased from OriGene (Rockville, MD, USA). Pooled human liver S9 fraction (HLS9) and pooled HlungS9 were products from XenoTech LLC (Kansas City, KS, USA). The demographic information about the tissue donors is summarized in Table S1. The ActivX™ desthiobiotin-FP probe and Pierce™ streptavidin agarose resin were obtained from Thermo Scientific (Rockford, IL, USA). Bis-(p-nitrophenyl) phosphate (BNPP), dithiolthreitol, iodoacetamide, urea, ammonium bicarbonate, and Tris-base (Trizma base) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tris-HCl was purchased from Fisher Scientific (Fair Lawn, NJ, USA). The MES buffer (0.2 M, pH 5.5) was purchased from Alfa Aesar (Ward Hill, MA, USA). All other chemicals and reagents were of analytical grade and commercially available.

Li J., Liu S., Shi J, & Zhu H.J. (2021). Activation of Tenofovir Alafenamide and Sofosbuvir in the Human Lung and Its Implications in the Development of Nucleoside/Nucleotide Prodrugs for Treating SARS-CoV-2 Pulmonary Infection. Pharmaceutics, 13(10), 1656.

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