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Bromophenol blue

Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, India, Italy, Canada, Australia, France

Bromophenol blue is a pH indicator dye commonly used in biochemistry and molecular biology applications. It is a chemical compound with the formula C₁₉H₁₀Br₄O₅S. Bromophenol blue exhibits a range of color changes across different pH levels, making it useful for monitoring the pH of solutions during various experiments and procedures.

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232 protocols using bromophenol blue

1

Vinblastine-Induced Protein Dephosphorylation

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When cells treated with vinblastine were harvested, the pellets were separated in two: one where total proteins were extracted in the presence of phosphatase inhibitors (10 mM NaF, 1 mM Na3VO4, 1 mM phenylmethanesulfonylfluoride, Sigma-Aldrich), and one where phosphatase inhibitors were omitted. The latter extracts were submitted to dephosphorylation with 200 units of λ-phosphatase (#P0753S, Biolabs, Ipswich, MA, USA) for 2 h at 37 °C. The reaction was stopped by 4% SDS, 125 mM tris pH = 6.8, 20% glycerol (Sigma-Aldrich), 0.002% (w/v) bromophenol blue (Sigma-Aldrich).
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2

Denaturing SDS-PAGE of B. pertussis Lysate

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An amount of 10 μg B. pertussis B1917 lysate (see sample preparation) was incubated with 4 × reducing sample buffer (250 mM Tris (Thermo Fisher, Merck, Netherlands), 8% SDS (Sigma Aldrich, Germany), 400 mM DTT (Sigma Aldrich, Germany), 40% Glycerol (Sigma Aldrich, Germany), and 0.04% bromophenol blue (Sigma Aldrich, Germany) 10 min at 100°C. Sample was loaded on 10% NuPAGE bis tris 1.0 mm precast gel (Thermo Fisher, Invitrogen, Netherlands) and proteins were separated with MES running buffer (Thermo Fisher, Invitrogen, Netherlands), 200 Volt (V) for 45 min in Xcell surelock minicell system (Thermo Fisher, Invitrogen, Netherlands). Gel was either stained with Coomassie (Imperial protein stain, Thermo Fisher, Netherlands) or used for Western blot.
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3

Synthesis of Linear-Dendritic Copolymers

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dl-Tyrosine (99+%),
2,2′-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS,
98+%), 1-hydroxybenzotriazole (HOBT, 95+%), bromophenol blue (95%),
rhodamine B base (97%), methyl orange (95+%), and p-nitrophenyl palmitate (all from Sigma-Aldrich, St. Louis MO) were
used as received. Chlorobenzene (99+%, Acros) was degassed with argon
and stored over 4 Å molecular sieves. Laccase was produced and
used as previously described.19 All linear–dendritic
and dendritic–linear–dendritic copolymers were synthesized
by the coupling of preformed reactive fragments20 (link) or via living ring-opening anionic polymerization of ethylene
oxide initiated by third-generation benzyl ether dendrons.21 (link) All linear–linear and linear–hyperbranched
copolymers were produced by controlled radical polymerization (ATRP).
Details are provided in the Supporting Information.
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4

Oligonucleotide Synthesis and Purification Protocols

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All synthetic oligonucleotides were purchased from Eurofins. PCR primers were desalted, DNAzymes were purified by Eurofins proprietary High Purity Salt Free reverse phase cartridge purification system and short RNA substrates were purified by HPLC. RNase-free ultrapure water was produced using a Milli-Q Advantage A10 Water Purification System equipped with a BioPak® Polisher from Merck Millipore. Phusion High-Fidelity PCR kit and TranscriptAid T7 High Yield Transcription kit were from Life Technologies. 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), sodium chloride, manganese chloride tetrahydrate, formamide, bromophenol blue, 40% 19:1 acrylamide/bis-acrylamide, N,N,N′,N′-tetramethylethylenediamine, ammonium persulfate, urea, agarose, ethidium bromide and phenol:chloroform:isoamyl alcohol 25:24:1 pH 8.0 was purchased from Sigma-Aldrich. Ethylenediaminetetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) were from Scharlau. Boric acid was from VWR. Klorrent bleach was from Nilfisk. The nucleic acid dyes SYTO 61, PO-PRO 1, DRAQ5, Hoechst 33258, DAPI, Propidium iodide and PicoGreen were all from Life Technologies. GelRed was from Biotium. ethidium bromide was from Sigma.
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5

Resveratrol-based Biochemical Assay

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Resveratrol, protease lytic buffer, and bromophenol blue were purchased from Sigma (MO, USA). Toluidine blue, paraformaldehyde, glycerol, 2,3,5-tritriphenyl-tetrazolium chloride (TTC), sodium hydroxide, potassium chloride, hydrogen peroxide, isoflurane, phosphate buffered solution, xylene and 3,3′-diaminobenzidine (DAB) were bought from Kangchen Biotechnology (Shanghai, China).
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6

Antioxidant and Cytotoxicity Assays

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Standards of ascorbic acid, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), potassium persulphate, 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), thiazolyl blue tetrazolium blue (MTT), bromophenol blue, and Tris–HCl were purchased from Sigma–Aldrich Chemical Co. (St. Louis, MO, USA). β-d-(+)-Glucose was purchased from ChromaDex (Irvine, CA, USA). Dulbecco’s Modified Eagle Medium (DMEM, high glucose + GlutaMAX), fetal bovine serum (FBS), Dulbecco’s phosphate-buffered saline (DPBS) were provided by Gibco (Life Technologies, Paisley, UK). Sodium dodecyl sulphate (SDS) was purchased from BioShop (Burlington, ON, Canada), Coomassie brilliant blue was from Fluka (Buchs, Switzerland), and dithiothreitol (DTT) from Serva (Heidelberg, Germany). Redistilled phenol, sulphuric acid, isoamyl alcohol, and solvents were provided by POCH (Gliwice, Poland). All the chemicals used were of analytical grade.
Assessment of biological activity and determination of total phenolic content was conducted on 96-well transparent microplates (Nunclon, Nunc, Roskilde, Denmark) using an Elisa Reader Infinite Pro 200F (Tecan Group Ltd., Männedorf, Switzerland). Total sugar, glucans, and protein content were determined using a UV-VIS Evolution 300 spectrophotometer (Thermo Scientific, Lafayette, CO, USA).
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7

Characterization of Xylanase Activity

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The p-nitrophenyl-β-d-xylopyranoside (pNPX), p-nitrophenyl-β-d-glucopyranoside (pNPG), beech wood xylan, N-ethylmaleimide (NEM), iodoacetate, phenyl methylsulfonyl fluoride (PMSF), diethyl-pyrocarbonate (DEPC), 1 ethyl-3-(3 dimethyl aminopropyl) carbidiimide (EDAC), 2-4-6 trinitrobenzenesulfonic acid (TNBS), 5-bromosuccinimide (NBS), N-acetylimidazole (NAI), citraconic anhydride, acetic anhydride, phenyl glyoxal, HEPES and MES, QAE-Sephadex A-50, Sephacryl-200 Coomassie Brillient Blue G-250, Bromophenol Blue were obtained from Sigma-Aldrich, St. Louis, USA. SDS-PAGE markers were purchased obtained from Invitrogen. All other chemicals were commercially sourced and used without further purification.
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8

Extraction and Characterization of Fish Skin Collagen

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Bovine face pieces were purchased from a local tannery. Scyliorhinus canicula fish skin waste was collected from the fish market, Monastir, Tunisia, and stored at −20°C until used. Sodium hydroxide (NaOH), sodium chloride (NaCl), and butanol (99%) were purchased from Loba Chemie, and acetic acid was purchased from Merck India Ltd. Acrylamide, bis-Acrylamide, Tris-HCl, SDS (sodium dodecyl sulfate), ammonium persulfate, TEMED (tetraacetylethylenediamine), glycerol, β-mercaptoethanol, bromophenol blue, and Coomassie R-250 were purchased from Sigma-Aldrich.
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9

Insulin Signaling Pathway Analysis

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RPMI-1640 culture medium and Fetal Bovine Serum were purchased from Gibco-Thermo Fisher Scientific. RPMI-1640 medium without phenol red, sodium chloride, potassium chloride, disodium phosphate, monopotassium phosphate, Nonidet 40, SDS, Na3VO4, PMSF, NaF, cOmplete cocktail 25x, crystal violet, EDTA, methanol, glycine, Tris-Base, NaOH, glutaraldehyde, acetic acid, beta-mercaptoethanol, bromophenol blue, glycerol, ammonium persulfate, TEMED, Tween-20, 1,1-Dimethylbiguanide hydrochloride, wortmannin, genistein were from Sigma Aldrich. 30% Acrylamide/Bisacrylamide Solution and Quick Start Bradford Protein Assay Dye Reagent from Biorad. Bortezomib was from Sandoz. Fast-acting recombinant human insulin (100 UI/ml) of PiSA. Recombinant human TNF-α of R&D Systems. Antibodies against IR-β (sc-81465), pIR-β (Tyr 1162–1163) (sc-25103), Akt-1 (sc-1618-R), pAkt (Ser 473) (sc-81433), p70S6K (sc-8418), pp70S6K (Thr 389) (sc-8416), Erk (sc-271269), pp38 (Tyr 182) (sc-166182), p38 (sc-7972), p65 (sc-372) and β-Actin (sc-47778) were from Santa Cruz Biotechnology. Antibody to pp65 (Ser 536) (93H1) and pErk (Thr 202-Tyr 204) (4370) of Cell Signaling Technology. Antibody to IKB-α (610690) of BD Transduction Laboratory. Anti-Mouse HRP and anti-Rabbit HRP secondary antibodies were of Invitrogen-Thermo Fisher Scientific. Oligos for RT-PCR were synthesized by Integrated DNA Technologies (IDT).
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10

Apoptosis Induction and Measurement

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TRAIL was purchased from Biomol (Plymouth Meeting, PA, USA). Propidium iodide (PI), Crystal violet, Bromophenol blue, DMSO, Tween 20, EDTA, DTT, Sucrose, HEPES, Tris and Glycine were purchased from Sigma Aldrich (St. Louis, MO, USA). Pan-caspase inhibitor ZVAD-fmk was obtained from Alexis Biochemicals, Lausen, Switzerland. Transfection reagents, Superfect and Dharmafect, were procured from Qiagen (Hilden, Germany) and Thermo Scientific (Waltham, MA, USA), respectively. CM-H2DCFDA and fluorogenic substrates of caspase-8, -9 and -3 were purchased from Invitrogen (Eugene, Oregon, USA). FeTPPS (5,10,15,20-tatrakis(4-sulfonatophenyl)porphyrinato iron (III) chloride) was purchased from Merck (Darmstadt, Germany) and ONOO was obtained from Cayman Chemical (Ann Arbor, MI, USA). Coomassie Blue protein concentration reagent was obtained from Pierce Biotechnology (Rockford, IL, USA). 10X phosphate buffered saline (PBS), 10X SDS, Tris-HCl buffer (pH 7.4) were purchased from NUMI Media Preparation Facility (National University of Singapore).
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