2 4 morpholino ethanesulfonic acid mes

Manufactured by Merck Group

Sourced in Belgium, United States

2-(4-Morpholino) ethanesulfonic acid (MES) is a chemical compound commonly used as a buffer in biological and biochemical applications. It maintains a stable pH range between 5.5 and 6.7, making it suitable for a variety of experiments and assays.

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6 protocols using 2 4 morpholino ethanesulfonic acid mes

Fabrication and Characterization of Magnetic Immunosensor

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Multiwalled carbon nanotubes (MWCNTs, SN2302, purity>95%) were purchased from Nanomaterial Store (Fremont, CA, https://www.nanomaterialstore.com/), FeCl₂·4H₂O (purity>99%) was purchased from Acros Organics BVBA (Geel, Belgium, http://www.acros.com/), FeCl₃·6H₂O, N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide (sulfo-NHS), 2-(4-Morpholino) ethanesulfonic acid (MES), streptavidin, sucrose, Tween 20, bovine serum albumin (BSA), Hexadecyltrimethylammonium bromide (CTAB) and phosphate buffer saline (0.01 M, pH 7.4) were purchased from Sigma-Aldrich (St. Louis, MO, http://www.sigmaaldrich.com/). Glass ﬁbers (GFCP000800), cellulose ﬁber (CFSP001700), nitrocellulose membranes (HF090MC100, HFB18004 and HFB24004) and laminated cards (HF000MC100) were purchased from Millipore (Billerica, MA, http://www.merckmillipore.com/). CA 19-9 protein, Mouse monoclonal CA 19-9 antibody 10-CA19A (Ab₁) and 10-CA19B (Ab₂) were purchased from Fitzgerald Industries International (Acton, MA, https://www.fitzgerald-fii.com/). Goat anti-mouse IgG (Ab₃) were purchased from ThermoFisher Scientific (Rockford, IL, https://www.thermofisher.com/).
All the chemicals used in this study were analytical reagent grade. Solutions were prepared with ultrapure (Z18 MΩ) water from Millipore Milli-Q water puriﬁcation system (Billerica, MA, http://www.merckmillipore.com).

Huang Y., Wen Y., Baryeh K., Takalkar S., Lund M., Zhang X, & Liu G. (2017). Lateral flow assay for carbohydrate antigen 19-9 in whole blood by using magnetized carbon nanotubes. Mikrochimica acta, 184(11), 4287-4294.

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Polycarbonate Surface Functionalization

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Polycarbonate (PC, granular, 3 mm nominal size), (3-Aminopropyl)triethoxysilane (APTES), 99%, succinic anhydride, ≥99%, N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), ≥98%, N-N-Diisopropylethylamine (EDIPA), 99.5%, MES hydrate, ≥99.5%, dodecylethyldimethylammonium bromide (DDAB), ≥98%, and 6-aminofluorescein, ≥95% were purchased from Sigma-Aldrich (St. Louis, MO, USA). PMSQ (Tospearl 120, dimeter of ~2 µm) were purchased from Momentive, Waterford, NY, USA. Silica nanoparticles (Aerosil OX 50, 40 nm, Evonik, Hanau, Germany). Methanol, ethanol, acetonitrile (ACN), toluene, and tetrahydrofuran (THF) of analytical grade with purity ≥99%, as well as ultra-pure deionized water (ULS/MS grade) were used as received without further purification. 2-(4-Morpholino)ethanesulfonic acid (MES) was purchased from Sigma-Aldrich Chemicals. Buffer 0.05 M was prepared by dissolving the appropriate amount of MES in deionized water.

Mani K.A., Yaakov N., Itzhaik Alkotzer Y., Zelikman E, & Mechrez G. (2018). A Robust Fabrication Method for Amphiphilic Janus Particles via Immobilization on Polycarbonate Microspheres. Polymers, 10(8), 900.

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Enzymatic Extraction of Polysaccharides from P. eryngii

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Fresh P. eryngii was obtained from Xinxiang Kanghong Edible Mushrooms Co., Ltd. Commercial wheat flour with 12.0% moisture, 1.5% fat, and 9.0% protein was obtained from COFCO Co., Ltd. Salt was purchased from the local market. High temperature‐resistant alpha‐amylase solution (30 U/mg) and glycosylase solution (100 U/mg) were purchased from Shanghai Ruji Bio‐Technology Co., Ltd. Alkaline protease solution (100 U/mg) was purchased from Shanghai Lanji Bio‐Technology Co., Ltd.; 2‐(4‐Morpholino)ethanesulfonic acid (MES) and tris(hydroxymethyl)aminomethane (TRIS) were bought from Sigma‐Aldrich Chemical Co. All other chemicals used in experiments were of analytical grades.

Nie Y., Zhang P., Deng C., Xu L., Yu M., Yang W., Zhao R, & Li B. (2019). Effects of Pleurotus eryngii (mushroom) powder and soluble polysaccharide addition on the rheological and microstructural properties of dough. Food Science & Nutrition, 7(6), 2113-2122.

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Antibody Validation for UPR Signaling

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Polyclonal antibodies for phosphorylated eIF2α (Ser51) and TBP (TATA-binding protein), as well as monoclonal antibodies for β-actin (clone 13E5) and GAPDH (clone 14C10), were purchased from Cell Signaling Technology (Danvers, MA, USA). Polyclonal antibodies for ATF6α and ATF4 (CREB2) were from Santa Cruz Biotechnology (Dallas, TX, USA). The antibody for ATF3 was purchased from GeneTex (Irvine, CA, USA), and the antibody for phosphorylated IRE1α (Ser724) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), N-(2-hydroxyethyl)-piperazine-N′-3-propanesulfonic acid (EPPS), 2-(4-morpholino)-ethanesulfonic acid (MES), and the protease inhibitor cocktail were from Sigma-Aldrich (St Louis, MO, USA) and Thermo Fisher Scientific (Waltham, MA, USA). The GPR4 inhibitor, 2-Ethyl-3-(4-((E)-3-(4-isopropyl-piperazin-1-yl)-propenyl)-benzyl)-5,7-dimethyl-3H-imidazo(4,5-b)pyridine, was purchased from Dalton Pharma Services (Toronto, ON, Canada).

Dong L., Krewson E.A, & Yang L.V. (2017). Acidosis Activates Endoplasmic Reticulum Stress Pathways through GPR4 in Human Vascular Endothelial Cells. International Journal of Molecular Sciences, 18(2), 278.

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Fluorescent Bead Conjugation Protocol

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Fluorescent beads were provided by the BioNano Health Guard Research Center (Daejeon, Korea). The surface of the fluorescent beads (diameter: 450 nm) was activated with 3 mM 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC; cat #77149, Pierce) and 3 mM N-hydroxysuccinimide (NHS; cat #56485, Sigma) in 50 mM 2-(4-morpholino)ethanesulfonic acid (MES; cat #2933, Sigma) buffer for 1 h. Activated fluorescent beads were centrifuged at 15,000 rpm for 15 min. After removing the supernatant, the fluorescent beads were mixed with 125 μg/mL mouse anti-human IgG and mouse anti-human IgM (Thermo Fisher scientific, USA) for 2 h. Next, 1/10 volume of 20% weight/volume skim milk (cat #232100, Gibco) was added, along with 1/10 volume of the second blocking solution (cat #ABF2BS, Absology, Korea), and incubated for 30 min at room temperature. The fluorescent beads were washed three times with storage buffer (cat #IBFSB, Absology), centrifuged, and the supernatant then removed. Pellets were resuspended in MES buffer, and the concentration of dAb-conjugated fluorescent beads was determined using a UV-1800 spectrometer (Shimadzu). Fluorescent beads at a concentration of 0.2% weight/volume in 1.5 μL conjugate buffer (cat #ABCB; Absology) were loaded onto the dAb deposition zone of the bottom substrate (Fig. 1E green box) using a nanoliter dispenser (Musashi, Japan) and then dried.

Lee J.H., Bae P.K., Kim H., Song Y.J., Yi S.Y., Kwon J., Seo J.S., Lee J.M., Jo H.S., Park S.M., Park H.S., Shin K.S., Chung S, & Shin Y.B. (2021). A rapid quantitative on-site coronavirus disease 19 serological test. Biosensors & Bioelectronics, 191, 113406.

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Herceptin-Conjugated PEGylated Nanobeads

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Herceptin molecules were covalently bound to the PEGylated NBs (NBs-Blank) by linking the free amino groups of Herceptin and the carboxyl groups of DSPE-PEG2000 on the NBs. Briefly, 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC, Sigma-Aldrich, St. Louis, Missouri) was mixed with N-hydroxysuccinamide (NHS, Sigma-Aldrich, St. Louis, Missouri) using an EDC:NHS:DSPE-PEG2000 molar ratio of 30:30:3 in a 2-(4-morpholino)ethanesulfonic acid (MES, Sigma-Aldrich, St. Louis, Missouri) solution (pH 5.5) for 30 min at room temperature. Then, the suspension was removed and centrifuged (1000 rpm, 5 min) three times to remove excess EDC and NHS. Herceptin (Hoffman La Roche, 1 mg/mL) was then added with a Herceptin/DSPE-PEG2000 molar ratio of 1:30 and incubated at 4 °C for 8 h (NBs-Her). Finally, the upper layer of the suspension was collected and washed (1000 rpm, 5 min) three times to remove the excess free Herceptin and stored at 4 °C.

Jiang Q., Hao S., Xiao X., Yao J., Ou B., Zhao Z., Liu F., Pan X., Luo B, & Zhi H. (2015). Production and characterization of a novel long-acting Herceptin-targeted nanobubble contrast agent specific for Her-2-positive breast cancers. Breast Cancer (Tokyo, Japan), 23(3), 445-455.

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