Five healthy and five CF airway mucus samples were ultracentrifuged at 32,000 rpm at 4°C for 1 hour to separate sol phase and gel phase. The sol phase was collected and incubated with 10% (v/v) of the treatment [PBS for healthy and CF control, catalase (100 U/ml), 1 N HCl] at 37°C for 30 min. The samples were then incubated with 1 mM carboxy-H2DCFDA (Life Technologies) and esterase (10 U/ml) (Sigma-Aldrich) at 37°C for 15 min. Serial dilutions of 10 mM hydrogen peroxide (Sigma-Aldrich) in PBS were used as standards. The plate was read at 495-nm excitation/527-nm emission. A standard curve was plotted from the average relative fluorescence unit (RFU) representing the concentrations of the standards. The concentrations of ROS in samples were calculated against the standard curve. Stock solutions of hydrogen peroxide were calibrated by measuring the absorbance at 240 nm. Dividing this absorbance by 43.6 gives the stock concentration in molar units.
Esterase
Manufactured by Merck Group
Sourced in United States
Esterase is a type of enzyme that catalyzes the hydrolysis of ester bonds, breaking down esters into an alcohol and a carboxylic acid. It plays a role in various biological processes and can be used in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Sodium cholate, by Merck Group (2 mentions)
Hexane, by Merck Group (2 mentions)
Dimethylformamide, by Merck Group (2 mentions)
Acetone, by Merck Group (2 mentions)
Porcine pepsin, by Merck Group (2 mentions)
Anhydrous potassium carbonate, by Merck Group (2 mentions)
Thionyl chloride, by Merck Group (2 mentions)
Ammonia solution, by Merck Group (2 mentions)
Sodium bicarbonate, by Merck Group (2 mentions)
Dialysis tubing, by Merck Group (2 mentions)
Porcine pancreatin, by Merck Group (2 mentions)
Silica gel, by Merck Group (2 mentions)
Carboxy h2dcfda, by Thermo Fisher Scientific (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Griess reagent kit, by Thermo Fisher Scientific (1 mentions)
13 protocols using esterase
1
Quantification of Airway ROS Levels
2
Zirconium Carbide Powder Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bulk ZrC powder was purchased from a commercial supplier (Smart‐Elements, Austria) and stored in a dark argon glovebox. Isopropanol (IPA) and ethanol (99.5%, anhydrous) were purchased from Aladdin Reagents (Shanghai, China). PEG‐NH2 was purchased from Shanghai Yare Biotech, Inc. (China). Esterase was purchased from Sigma Aldrich. The AO/PI Assay Kit was obtained from Logos Biosystems (Korea). PBS (pH 7.4), FBS, RPMI‐1640 medium, trypsin‐EDTA, and penicillin/streptomycin were purchased from Gibco Life Technologies. All other chemicals used in this study were of analytical reagent grade and used without further purification. Ultrapure water (18.25 MΩ cm−1, 25 °C) was used to prepare all of the solutions.
3
Cellular Antioxidant and Anti-Inflammatory Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2,2′-Azobis (2-methylpropionamidine) dihydrochloride (AAPH), 2-methyl-1,4-naphthoquinone (menadione), iron (II) sulfate (FeSO4), hydrogen peroxide (H2O2), LPS (Escherichia coli serotype O111:B4), 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB), N-acetyl-L-cysteine (NAC), esterase, ConA, mitomycin C, dexamethasone, and BCA protein reagent were obtained from Sigma–Aldrich (St. Louis, MO, USA). The Griess reagent kit and 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Molecular Probes (Eugene, OR, USA). Dulbecco’s Modified Eagle’s medium (DMEM), Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), and streptomycin-penicillin were purchased from GIBCO (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The ONE-Glo™ luciferase assay system and FuGENE® HD transfection reagent were purchased from Promega (Madison, WI, USA). Hispolon and primary antibodies against phospho-IκBα, IκBα, NF-κB p65, JAK1, phospho-JAK1, STAT3, and transcription factor (TF) IIB were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Primary antibodies against phospho-STAT3 and α-tubulin and anti-rabbit secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
4
Oxidative Stress Assay Protocol
5
Nanomaterial Synthesis and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Poly(styrene-alt-maleic acid)sodium salt, 13 wt. % solution in water (PSMA, Mw = 350000) (Sigma-Aldrich), poly(4-styrene sulfonic acid-co-maleic acid) sodium salt (Mw = 20000) (Aldrich), poly(maleic anhydrate-alt-1-octadence) (Mw = 30000–50000) (Aldrich), succinic acid (Sigma), maleic acid (99.5%) (Chem Service), quantum dot (Life), hydrogen tetrachloroaurate(III) trihydrate (HAuCl4, 99.99%) (Alfa Aesar), Silver nitrate (AgNO3) (Fisher), Copper(II) chloride dihydrate (CuCl2·2H2O) (Riedel-de Haen), Iron(II) chloride tetradhydrate (FeCl2·4H2O) (J. T. Baker), Hydrazine monohydrate (N2H4·H2O, 98%) (Alfa Aesar), titanium(IV) isopropoxide (97%) (Sigma-Aldrich), tetraethyl orthosilicate (TEOS, 98%) (Sigma-Aldrich), sodium hydroxide (NaOH, min 99%) (Fullin), methylene blue (MB, high purity) (Alfa Aesar), Chlorin e6 (Ce6) (Frontier), doxorubicin hydrochloride (DOX) (Sigma-Aldrich), esterase (Sigma), 3,3′,5,5′-Tetramethylbenzidine (TMB, ≥99%) (Sigma-Aldrich) were purchased for use without purification.
6
Synthesis of Biocompatible Polymer Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2-Hydroxyethyl 2′-methyl-2′-bromopro-pionate (HMB) was purchased from Changzhou Yipin Tang Chemical Co. LTD. Chloroauric acid (HAuCl4), Sn (Oct)2, ε-caprolactone (ε-CL) and 2-(2-methoxyethoxy) ethyl methacrylate (MEO2MA) were purchased from Aladdin reagent (Shanghai, China). Calcein-AM, propidium iodide (PI), esterase, methyl thiazolyl tetrazolium (MTT), copper bromide (CuBr) and N, N, N′, N′, N″-pentamethyldiethylenetriamine (PMDETA) were purchased from Sigma-Aldrich (St. Louis, Missouri).
7
Disodium Cromoglycate Synthesis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Disodium cromoglycate (5′DSCG) (98% purity) was obtained from TCI Chemicals (Philadelphia, PA). Tryptone, yeast extract, sodium chloride, Tris base, urea, lauryldimethylamine N-oxide and poly(ethylene) glycol (PEG8000) were purchased from Fisher Scientific (Fair Lawn, NJ) and were used to make Luria Bertani media and buffers. All aqueous solutions were dissolved in deionized water with resistivity greater than 18.2 MΩ cm or deuterium oxide (D2O, 99.9%, Cambridge Isotope Laboratories, Inc., Andover, MA). Casamino acids (Amresco, Solon, OH), peptone (Bio Basic, Amherst, NY) and casitone (BD Biosciences, MD) were also used. l -amino acids (l -glutamic acid, l -alanine, l -arginine), polyvinylpyrrolidone (PVP, MW ∼ 40 000), polyvinylalcohol (PVA, MW ∼ 9000–10 000), poly-acrylamide (PAAm, MW ∼ 9000–10 000), poly(ethylene) glycol (PEG4000), and proteins (lectin A, esterase, lipase, bovine serum albumin and trypsin) were purchased from Sigma Aldrich (St. Louis, MO). Pseudomonas aeruginosa strain PA1244N3 (pPAC46) was obtained from Dr Castric and Dr Horzempa.28,29 (link)
8
Synthesis and Characterization of MTDZ Prodrugs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dextran (MW 70,000), dextranase (Penicillium sp.), esterase, 2,4-dinitrosalicylicacid (DNS), succinic anhydride, carbonyldiimidazole (CDI), and MTDZ were obtained from Sigma-Aldrich Chemical Co. (St Louis, MO, USA). Dimethyl sulfoxide (DMSO)-d6 was obtained from Cambridge Isotope Laboratories (Andover, MA, USA). All reagents and solvents for high-performance liquid chromatography (HPLC) were obtained from Merck (Darmstadt, Germany). All other chemicals used were reagent grade, commercially available products. Infrared (IR) spectra were recorded on a Fourier transform-infrared (FT-IR) spectrophotometer (Varian, Palo Alto, CA, USA). 1H-nuclear magnetic resonance (NMR) spectra were obtained using a Varian AS 500 spectrometer, and the chemical shifts were recorded in parts per million (ppm) downfield from tetramethylsilane. Small molecular colon-specific prodrugs of MTDZ, MTDZ sulfate, and NMG were synthesized as described in previous studies.8 (link),9 (link) Structures of the prodrugs are shown in Figure S1 .
9
Esterase-Mediated FTs Degradation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The FTs solution was incubated with esterase (Sigma) and hydrogen peroxide for different lengths of time, the degradation of FTs was then monitored by measuring the metal-to-ligand charge transfer band (MLCT) using UV-Vis spectrophotometer (Agilent 8453).
10
Comparative Evaluation of Lipase Preparations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Commercially available lipases were used as follows: (i) Immobilized lipases:
Candida antarctica lipase type B immobilized on acrylic resin (CAL-B, 7,300.0 U/g), Thermomyces lanuginosus lipase immobilized on immobead-150 (TLL, 250.0 U/g), Rhizopus oryzae lipase immobilized on immobead-150 (ROL, 340.0 U/g), Pseudomonas fluorescens lipase immobilized on immobead-150 (PFL, ≥ 600 U/g), Pseudomonas fluorescens lipase immobilized in Sol-Gel (AK, ≥ 30 U/g), Pseudomonas fluorescens lipase immobilized in Sol-Gel on pumice (AK, ≥ 8.0 U/g) and Amano lipase PS from Burkholderia cepacia immobilized on diatomaceous earth (PS-IM, ≥ 500 U/g) were acquired from Sigma-Aldrich®, Rhizomucor miehei lipase immobilized in anionic resin (RML, 150.0 U/g) was purchased from Novozymes ® . (ii) Unsupported lipase preparations: Pseudomonas fluorescens lipase (AK, 22,100.0 U/g), Penicillium camemberti lipase (G, 50.0 U/g), lipase from Rhizopus niveus (RNL, ≥ 1.5 U/mg), Amano lipase from Mucor javanicus (MJL, ≥ 10,000 U/g) and lipase from Aspergillus niger (ANL, 200 U/g) were acquired from Sigma-Aldrich ® . Porcine pancreas lipase (PPL, 46.0 U/g solid), and Candida rugosa lipase (CRL, 1.4 U/g) were obtained from Sigma-Aldrich ® . (iii) Esterase, immobilized in Eupergit ® C from hog liver (~200 U/g) was obtained from Sigma-Aldrich ® .
Candida antarctica lipase type B immobilized on acrylic resin (CAL-B, 7,300.0 U/g), Thermomyces lanuginosus lipase immobilized on immobead-150 (TLL, 250.0 U/g), Rhizopus oryzae lipase immobilized on immobead-150 (ROL, 340.0 U/g), Pseudomonas fluorescens lipase immobilized on immobead-150 (PFL, ≥ 600 U/g), Pseudomonas fluorescens lipase immobilized in Sol-Gel (AK, ≥ 30 U/g), Pseudomonas fluorescens lipase immobilized in Sol-Gel on pumice (AK, ≥ 8.0 U/g) and Amano lipase PS from Burkholderia cepacia immobilized on diatomaceous earth (PS-IM, ≥ 500 U/g) were acquired from Sigma-Aldrich®, Rhizomucor miehei lipase immobilized in anionic resin (RML, 150.0 U/g) was purchased from Novozymes ® . (ii) Unsupported lipase preparations: Pseudomonas fluorescens lipase (AK, 22,100.0 U/g), Penicillium camemberti lipase (G, 50.0 U/g), lipase from Rhizopus niveus (RNL, ≥ 1.5 U/mg), Amano lipase from Mucor javanicus (MJL, ≥ 10,000 U/g) and lipase from Aspergillus niger (ANL, 200 U/g) were acquired from Sigma-Aldrich ® . Porcine pancreas lipase (PPL, 46.0 U/g solid), and Candida rugosa lipase (CRL, 1.4 U/g) were obtained from Sigma-Aldrich ® . (iii) Esterase, immobilized in Eupergit ® C from hog liver (~200 U/g) was obtained from Sigma-Aldrich ® .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!