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Tris base

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Tris base is a laboratory chemical compound commonly used as a buffer in biochemical and molecular biology applications. It is a white crystalline solid that provides a stable and physiologically compatible pH environment for various biological experiments and procedures.

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Lab products found in correlation

Glycine, by Bio-Rad (3 mentions) Nitrocellulose membrane, by Bio-Rad (3 mentions) Agarose gel, by Bio-Rad (3 mentions) Tris acetic acid edta tae, by Bio-Rad (3 mentions) Sodium dodecyl sulfate (sds), by Bio-Rad (3 mentions) Ammonium persulfate, by Bio-Rad (2 mentions) Bis acrylamide, by Bio-Rad (2 mentions) Ethidium bromide, by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Trolox, by Merck Group (2 mentions) Acetic acid, by RCI Labscan (2 mentions) Trichloroacetic acid, by RCI Labscan (2 mentions) Folin ciocalteu reagent, by Merck Group (2 mentions) Glacial acetic acid, by Thermo Fisher Scientific (1 mentions) Acrylamide, by Thermo Fisher Scientific (1 mentions) Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions) Ethylenediaminetetraacetic acid, by Bio-Rad (1 mentions) N n n n tetramethylethylenediamine, by Bio-Rad (1 mentions) Precision plus protein standard, by Bio-Rad (1 mentions) Urea, by Thermo Fisher Scientific (1 mentions)

13 protocols using tris base

1

Quantitative Milk Protein Analysis

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We obtained the materials from Sigma-Aldrich (Saint Louis, MO, USA): bovine BC, a-LA, b-LG (purity ≥ 98%) and IgG (≥95%) protein standards, human milk LF protein standard (>95%), 1,4-dithiothreitol (DTT), iodoacetamide, acetonitrile, formic acid, and thermolysin (Type X, E.C. No. 3.4.24.27). Tris base, ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), ammonium persulfate, N, N, N, N’-tetramethylethylenediamine (TEMED), Precision Plus ProteinTM standards, and colloidal Coomassie G-250 stain, all molecular biology grade, were acquired from Bio-Rad (Hercules, CA, USA). Acrylamide, N, N’-methylenebisAcrylamide, urea, 2-mercaptoethanol, glycine and bromophenol blue, and ultrapure Bioreagent grade, as well as isobutyl alcohol, glacial acetic acid, and sodium hydroxide, were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Thermo Fisher Scientific provided Invitrogen’s Qubit® Protein Assay kit. Zip Tips C18 were obtained from Millipore Sigma (Saint Louis, MO, USA). Deionized water, used in all experiments, was purified using a Milli-Q system from Millipore Merck (Darmstadt, Germany).
Muñoz-Salinas F., Andrade-Montemayor H.M., De la Torre-Carbot K., Duarte-Vázquez M.Á, & Silva-Jarquin J.C. (2022). Comparative Analysis of the Protein Composition of Goat Milk from French Alpine, Nubian, and Creole Breeds and Holstein Friesian Cow Milk: Implications for Early Infant Nutrition. Animals : an Open Access Journal from MDPI, 12(17), 2236.
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2

Western Blot Analysis Protocol

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Samples were separated by Novex NuPAGE SDS-PAGE system using 4-12% BisTris gels in MES running buffer according to manufacturer’s instructions. All steps were done at RT. Proteins were transferred to Amersham Hybond P 0.2 μm pore size PVDF membrane in a BioRad Mini Trans-Blot Cell at 25 V constant for 16 h in SDS-free transfer buffer (50 mM Tris base (Trizma), 384 mM glycine). Membranes were blocked in 5% (w/v) milk powder in PBS (blocking solution) for 15 min and incubated for 3 h with the primary antibody diluted in blocking solution at concentrations indicated in the Key Resource Table. Membranes were washed three times in PBS containing 0.2% Tween 20 (v/v) for 5 min and then incubated for 1 h at RT with the respective species-specific secondary antibody coupled to horseradish peroxidase. Membranes were washed three times in PBS containing 0.2% Tween 20 (v/v) for 10 min and incubated in ECL Plus Western Blotting Substrate (Thermo Scientific) for 5 min. Chemiluminescence was documented on a ChemiDoc MP (BioRad) system. All immunoblots were recorded with no saturated pixels.
Samwer M., Schneider M.W., Hoefler R., Schmalhorst P.S., Jude J.G., Zuber J, & Gerlich D.W. (2017). DNA Cross-Bridging Shapes a Single Nucleus from a Set of Mitotic Chromosomes. Cell, 170(5), 956-972.e23.
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3

HPV Multiplex Primer and Western Blot Analysis

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HPV multiplex primers, Agarose powder, Ethylenediaminetetraacetic acid (EDTA), 2- deoxyribonucleic acid (DNA) ladder and ethidium bromide were from Sigma-Aldrich (France). Tris-Base, glycine, sodium dodecyl sulfate, bis-acrylamide and nitrocellulose membrane, the protein ladder and the peroxidase conjugated secondary antibodies anti-rabbit and anti-mouse were purchased from Bio-Rad Inc (USA). Sodium chloride (NaCl), potassium chloride (KCl), Tween-20, protease inhibitor phenyl-methyl-sulfonyl fluoride (PMSF), 2-mercaptoethanol, methanol, glycerol, 1,4-Dithiothreitol (DTT), sodium fluoride (NaF), sodium azide (NaN3), Tris-Hydrochloride (tris-HCl) and sodium dodecyl sulfate (SDS, NaC12H25SO4) were from Sigma-Aldrich (USA). The primary antibody against lamin A/C was purchased from Transduction Lab (USA). Antibody against ß-tubulin was from Santa Cruz Biotechnology (CA, USA). The chemo-luminescence reagent “Super Signal West Dura Extended Duration Substrate” made by PIERCE was from Thermo Scientific (Rockford, IL USA) and was used on Western blot membranes for protein revelation after exposure to X-ray films [12 (link)].
Capo-chichi C.D., Aguida B., Chabi N.W., Acapko-Ezin J., Sossah-Hiffo J., Agossou V.K., Anagbla T., Zannou M., Houngbé F, & Sanni A. (2016). Diversity of high risk human papilloma viruses in women treated with antiretroviral and in healthy controls and discordance with cervical dysplasia in the South of Benin. Infectious Agents and Cancer, 11, 43.
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4

2D Gel Electrophoresis Proteomic Protocol

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Cy2, Cy3, and Cy5 were purchased from GE Healthcare. dimethylformamide was purchased from Aldrich. DTT, urea, agarose, glycerol, bromphenol blue, CHAPS, mineral oil, acrylamide, Bis, Tris base, glycine, SDS, iodoacetamide, ammonium persulfate, TEMED, Immobiline DryStrip gels (24 cm, pH 3–10), and Bio-Lyte solutions (pH 3–10) were purchased from Bio-Rad. Thiourea was purchased from Fluka (Buchs, Switzerland). Protease inhibitor mixture was purchased from Roche Applied Science. ACN and methanol were purchased from Fisher. TFA was purchased from Merck. Trypsin (sequencing grade) was purchased from Promega (Madison, WI). All buffers were prepared with Milli-Q water (Millipore, Bedford, MA).
Wang G., zhang Z., Yang M., Xu B., gao Q, & Yang X. (2016). Comparative proteomics analysis of human osteosarcoma by 2D DIGE with MALDI-TOF/TOF MS. Journal of Bone Oncology, 5(4), 147-152.
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5

Silymarin and Silibinin Cytotoxicity Assay

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Silymarin (SM, S0292), silibinin (SB, S0417), RPMI 1640 medium, methyl methanesulfonate (MMS), ethidium bromide, dimethyl sulfoxide (DMSO), NaCl, and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Low melting point agarose and normal melting point agarose were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Na2EDTA, Tris base, and Tris-HCl were purchased from Bio-Rad Laboratories (Hercules, CA, USA).
Fernandes Veloso Borges F., Ribeiro e Silva C., Moreira Goes W., Ribeiro Godoy F., Craveiro Franco F., Hollanda Véras J., Luiz Cardoso Bailão E.F., de Melo e Silva D., Gomes Cardoso C., Divino da Cruz A, & Chen-Chen L. (2018). Protective Effects of Silymarin and Silibinin against DNA Damage in Human Blood Cells. BioMed Research International, 2018, 6056948.
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6

Assessing Protein Interactions in Neurodegeneration

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Chemical reagents were purchased from the following sources: Mn chloride tetrahydrate (MnCl2·4H2O) from Fisher Scientific (Pittsburgh, PA); Cu chloride (CuCl2), calcium chloride (CaCl2), Dextran-70, hydroxyethyl piperazineethanesulfonic acid (HEPES), monoclonal anti-mouse β-actin antibody, 2-mercaptoethanol, phenylmethylsulfonyl fluoride (PMSF), polyacrylamide and tetramethyl-ethylenediamine (TEMED) from Sigma Chemicals (St Louis, MO); ultrapure nitric acid from VWR international (Chicago, IL); Fluor Alexa-488 conjugated secondary antibody from Life Technologies (Carlsbad, CA); protease inhibitor cocktail from Calbiochem (San Diego, CA); Tris base, glycine, sodium dodecyl sulfate (SDS), 2xLaemmli sample buffer, Triton X-100, and clarity Western ECL substrate from Bio-Rad (Hercules, CA); Aβ40 pure PTD human protein and Aβ40 human ELISA kit from Invitrogen (Waltham, MA); anti-RAGE antibody and anti-LRP1 antibody from Abcam (Cambridge, MA); Anti-Aβ40 antibody from Biolegend (San Diego, CA); rat LRP1/CD91 ELISA Kit and rat AGER/RAGE ELISA Kit from LifeSpan BioSciences (Seattle WA); Radioactive 14C-sucrose (specific activity: 495 mCi/mmol) from Moravek Biochemicals (Brea, CA); and Eco-lite-(+) scintillation cocktail from MP Biomedicals (Irvine, CA). All reagents were of analytical grade, HPLC grade, or the best available pharmaceutical grade.
Shen X., Xia L., Liu L., Jiang H., Shannahan J., Du Y, & Zheng W. (2020). Altered clearance of beta-amyloid from the cerebrospinal fluid following subchronic lead exposure in rats: Roles of RAGE and LRP1 in the choroid plexus. Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), 61, 126520.
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7

HSYA Isolation and Analysis in C. tinctorius

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HSYA was isolated and purified from the aqueous extract of C. tinctorius L. by macroporous resin-gel column chromatography, as described previously14 (link). The molecular weight of HSYA is 612. HSYA was analysed using a high-performance liquid chromatography system (Shimadzu, Kyoto, Japan). iTRAQ kits were purchased from Applied Biosystems (Waltham, MA, USA), the 2-D Quant Kit from GE Healthcare (Pittsburgh, PA, USA), sequence-grade modified trypsin from Promega (Madison, WI, USA), and human ox-LDL (BT-910) from Alfa Aesar (Heysham, Lancashire, UK). Aspirin was the product of Sigma-Aldrich (St. Louis, MO, USA). Bromophenol blue, tetramethylethylenediamine, Coomassie brilliant blue G-250, Bis, low-molecular weight marker, nitrocellulose membrane, Tris-base, and the Bradford method protein assay kit were purchased from Bio-Rad (Hercules, CA, USA). VDAC-2 polyclonal antibody was purchased from Cell Signaling Technology (Danvers, MA, USA), and GAPDH mouse monoclonal antibody was from Beyotime Biotechnology (Jiangsu, China). Other polyclonal and monoclonal antibodies and siRNA reagents were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), with the secondary antibodies obtained from LI-COR Biosciences (Lincoln, NE, USA). NO and MDA assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
Ye F., Wang J., Meng W., Qian J, & Jin M. (2017). Proteomic investigation of effects of hydroxysafflor yellow A in oxidized low-density lipoprotein-induced endothelial injury. Scientific Reports, 7, 17981.
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8

Western Blot Analysis of Lamin and Caspase-6

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Tris-Base, glycine, sodium dodecyl sulfate, bis-acrylamide, nitrocellulose membrane, were purchased from Bio-Rad. Inc. (USA). NaCl, KCl, Tween-20, protease inhibitor PMSF, 2-mercaptoethanol, DTT, methanol, ethanol, EDTA, glycerol, sodium azide, sodium fluoride. The primary antibodies made in rabbit against lamin A/C, lamin A and cleaved lamin A were from Transduction Lab (USA). The primary rabbit antibodies for simultaneous detection of procaspase-6 and caspase-6 were from Sigma-Aldrich (USA) and Cell signaling. Peroxidase (HRP)-conjugated secondary antibody (anti-rabbit) made in goat was from Bio-Rad Inc. (USA). A Super Signal West Dura Extended Duration Substrate made by PIERCE was purchased from Thermo Scientific (Rockford, IL USA). Caspase-6 specific inhibitor drug A6339 (N-Acetyl-Val-Glu-Ile-Asp-aldehyde, Synonym: Ac-VEID-CHO) was purchased from Sigma-Aldrich, USA.
Capo-chichi C.D., Cai K.Q, & Xu X.X. (2018). Overexpression and cytoplasmic localization of caspase-6 is associated with lamin A degradation in set of ovarian cancers. Biomarker Research, 6, 30.
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9

Comet Assay for Nanoparticle-Induced DNA Damage

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Nanoparticle treated cells were embedded in 0.5% low melting agarose (Sigma Aldrich) layer between 1.0% normal melting agarose (Sigma Aldrich) and 0.5% normal melting agarose. The cells were lysed with high salt and detergent concentrations (100 mM EDTA (Sigma Aldrich), 2.5 M NaCl (Sigma Aldrich), 10 mM tris base (Bio Rad), 1% Triton X-100 (Sigma Aldrich), adjusted to pH 10) for 1 h. DNA was allowed to unwind (1 mM EDTA, 10% DMSO, 300 mM NaOH (Sigma Aldrich), pH 13) for 20 min and then subjected to electrophoresis in the same solution as for unwinding (25 V, 300 mA) for 15 min. After electrophoresis, the alkalis in the gels were neutralized by rinsing the slides in a neutralization buffer (0.1 M Tris pH 7.5) for 5 min. The slides were treated with methanol for 10 minutes, stained with 45 μl of 20 μg/ml ethidium bromide solution and viewed under a fluorescent microscope (Olympus Optical Co. Ltd). 1000 cells were analyzed and % tail DNA was measured to evaluate the extent of DNA damage. Results were expressed as means of at least three replicates ± standard error.
Dubey A., Goswami M., Yadav K, & Chaudhary D. (2015). Oxidative Stress and Nano-Toxicity Induced by TiO2 and ZnO on WAG Cell Line. PLoS ONE, 10(5), e0127493.
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10

Antioxidant Evaluation of Compounds

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ABTS radical cation (2,2′-azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid), anthrone, DPPH (2,2-Diphenyl-1-picrylhydrazyl, (−)-epigallocatechin gallate (EGCG), gallic acid, sulforhodamine B (SRB), and trolox were obtained from Sigma Chemical (St. Louis, MO, USA). Folin–Ciocalteu reagent was purchased from Merck (Darmstadt, Germany). Dutasteride, finasteride, and minoxidil were from Wuhan W&Z Biotech (Wuhan, China). Agarose gel, Tris base, and 50X Tris/Acetic acid/EDTA (TAE) were from Bio-Rad Laboratories (Hercules, CA, USA). Follicle Dermal Papilla Cell Growth Medium Kit (cat no. C-26501) was from Promo Cell GmbH (Heidelberg, Germany). Antibiotic-antimycotic (100X; cat no.1 5240062), fetal bovine serum (FBS; cat no. 16000044), and Roswell Park Memorial Institute medium (RPMI-1640; cat no. 31800022) were from Gibco Life Technologies (Thermo Fisher Scientific, Waltham, MA, USA). Acetic acid, dimethyl sulphoxide (DMSO), trichloroAcetic acid, sulfuric acid, and other chemical substances were purchased from RCI Labscan (Bangkok, Thailand). All other chemicals were analytical-grade substances.
Ruksiriwanich W., Khantham C., Muangsanguan A., Phimolsiripol Y., Barba F.J., Sringarm K., Rachtanapun P., Jantanasakulwong K., Jantrawut P., Chittasupho C., Chutoprapat R., Boonpisuttinant K, & Sommano S.R. (2022). Guava (Psidium guajava L.) Leaf Extract as Bioactive Substances for Anti-Androgen and Antioxidant Activities. Plants, 11(24), 3514.
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