S2770

Manufactured by Merck Group

The S2770 is a laboratory instrument manufactured by Merck Group. It is designed to perform specific tasks in a laboratory setting. The core function of the S2770 is to accurately measure and analyze samples. For detailed technical specifications and intended use, please consult the product literature or contact a Merck sales representative.

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Lab products found in correlation

W3500, by Merck Group (3 mentions) Chemically defined lipid concentrate, by Thermo Fisher Scientific (1 mentions) Sevencompact ph meter, by Mettler Toledo (1 mentions) Vapro 5600 vapor pressure osmometer, by Wescor (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) P0500, by Merck Group (1 mentions) O7501, by Merck Group (1 mentions) M3128, by Merck Group (1 mentions) P9417, by Merck Group (1 mentions) Collagen type 1, by Merck Group (1 mentions) P5493, by Merck Group (1 mentions) H3537, by Merck Group (1 mentions) P35g 0 14 c, by MatTek (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)

6 protocols using s2770

Peptide-based Hydrogel Fabrication

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A commercially available peptide preparation in powder form was used as the source of the octapeptide gelator (FEFEFKFK, Phe-Glu-Phe-Glu-Phe-Lys-Phe-Lys). As part of this study we used peptide sourced from Cambridge Research Biochemicals (batch 32597) although we also verified the fabrication method using a second peptide source (Pepceuticals, UK). To form each precursor, a mass of between 7.5 and 18.75 mg peptide preparation was dissolved in 800 μL sterile water (W3500 Sigma), using a 3 min vortex step followed by centrifugation (3 min at 1000 rpm) and a 2 h incubation at 80 °C. After incubation, 0.5 M NaOH (S2770 Sigma) was added incrementally to the gels until optically clear. Gels were vortexed, buffered by addition of 100 μL 10× PBS (70011 Gibco), and incubated at 80 °C overnight. The resulting precursors could be stored at 4 °C until required.

Ashworth J.C., Thompson J.L., James J.R., Slater C.E., Pijuan-Galitó S., Lis-Slimak K., Holley R.J., Meade K.A., Thompson A., Arkill K.P., Tassieri M., Wright A.J., Farnie G, & Merry C.L. (2019). Peptide gels of fully-defined composition and mechanics for probing cell-cell and cell-matrix interactions in vitro. Matrix biology : journal of the International Society for Matrix Biology, 85-86, 15-33.

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Optimizing Pluripotent Cell Culture

Cited 1 time

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All pluripotent and reprogramming cell cultures were maintained at 37°C with 5% CO₂ and 5% O₂. Differentiation cultures were maintained at 5% CO₂ and atmospheric (∼21%) O₂. E8 medium was made in house as previously described (Burridge et al., 2015 , Chen et al., 2011 (link)). Other medium components tested were human Long R3 IGF1 (Sigma, 91590C), thiazovivin (LC Labs, T-9753), recombinant human TGF-β3 (Cell Guidance Systems, GFH109), sodium bicarbonate (Sigma, S5761), NEAA (Gibco, 11140050), Chemically Defined Lipid Concentrate (Gibco, 11905031), fatty acid-free BSA (GenDEPOT, A0100), and heparin sodium salt (Sigma H3149-250KU). The pH was adjusted with 1 N HCl (Sigma, H9892) or 1 N NaOH (Sigma, S2770) and measured at room temperature and atmospheric CO₂ using a SevenCompact pH meter (Mettler Toledo). Osmolarity was adjusted with sodium chloride (Sigma, S5886) or cell culture water (Corning, 25-055-CV) and measured with a Vapro 5600 vapor pressure osmometer (Wescor).

Kuo H.H., Gao X., DeKeyser J.M., Fetterman K.A., Pinheiro E.A., Weddle C.J., Fonoudi H., Orman M.V., Romero-Tejeda M., Jouni M., Blancard M., Magdy T., Epting C.L., George AL J.r, & Burridge P.W. (2020). Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem Cell Reports, 14(2), 256-270.

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Modulation of Type I Interferon Response by Fatty Acids

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To test the effects of free saturated and unsaturated fatty acids in modulating the type I interferon response, we prepared palmitate-BSA, palmitoleate-BSA, myristate-BSA, and sodium oleate-BSA conjugates. In brief, to prepare the BSA solution, 10% (w/v) fatty acid-free bovine albumin (A9418-5G, MilliporeSigma) was gradually added to ultrapure water at 52°C with gentle agitation until the BSA was fully dissolved. To prepare palmitate, palmitoleate, and myristate, palmitic acids (P0500, Sigma-Aldrich), palmitoleic acids (P9417, Sigma-Aldrich), and myristic acids (M3128, Sigma-Aldrich) were added to 0.1M NaOH (S2770, Sigma-Aldrich) and heated to 70°C until fully dissolved, yielding a concentration of 100 mM. Likewise, sodium oleate (O7501, Sigma-Aldrich) was added to ultrapure water at 55°C to yield a concentration of 100 mM. To conjugate fatty acids to BSA, the 100 mM stocks of palmitate, palmitoleate, myristate, and sodium oleate were added to 10% BSA at a ratio of 1:9 and heated at 55°C for 10 min. After conjugation, both the 10% (w/v) BSA solution and fatty acid-BSA solutions were filter-sterilized using a 0.45 μm PES filter (SLHVR33RS, Fisher Scientific), aliquoted, and stored at −20°C. Before use, solutions were warmed to 37°C. For final use, 200 μM of both fatty acid-BSA conjugates were used.

Heath B.R., Gong W., Taner H.F., Broses L., Okuyama K., Cheng W., Jin M., Fitzsimonds Z.R., Manousidaki A., Wu Y., Zhang S., Wen H., Chinn S.B., Bartee E., Xie Y., Moon J.J, & Lei Y.L. (2023). Saturated fatty acids dampen the immunogenicity of cancer by suppressing STING. Cell reports, 42(4), 112303.

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Fabrication of Collagen-Based Hydrogels

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Type-I collagen (Elastin Products Company, C857, Owensville, MO) was reconstituted in 0.02 M acetic acid at 3.75 mg/mL. CMA was synthesized from the solubilized Type-I collagen in the method outlined in Gaudet et al.^{35 (link)} and was also reconstituted in 0.02 M acetic acid at 3.75 mg/mL. For free-form fabrication, unless otherwise stated, CMA was buffered on ice according to the following formula: 20 μL 1 M HEPES (H3537, Sigma-Aldrich, St. Louis, MO), 134 μL 0.15 N NaOH (S2770, Sigma-Aldrich), 100 μL 10X PBS (P5493, Sigma-Aldrich), 59 μL 1X PBS (860454, Thermo Fisher Scientific, Waltham, MA), 10 μL 10% Irgacure (I2959, a gift from Ciba Specialty Chemicals) solubilized in neat methanol, and 677 μL CMA. Hydrogels were always prepared to a final concentration of 2.5 mg/mL and pH 7 unless otherwise specified. Buffered CMA was plated in 14 mm glass-bottomed MatTek dishes (P35G-0-14-C, MatTek Corporation, Ashland, MA) and self-assembled for one hour (unless otherwise stated) at 37 °C to create CMA hydrogels. All samples exposed to UV were plated in MatTek dishes.

Drzewiecki K.E., Malavade J.N., Ahmed I., Lowe C.J, & Shreiber D.I. (2017). A thermoreversible, photocrosslinkable collagen bio-ink for free-form fabrication of scaffolds for regenerative medicine. Technology, 5(4), 185-195.

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Hydrogel Precursor Synthesis Protocol

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The precursor and gel preparation method was followed as previously published⁵⁷. A commercially available peptide preparation in powder form was used as the source of the octapeptide gelator (Pepceuticals UK, FEFEFKFK, Phe-Glu-Phe-Glu-Phe-Lys-Phe-Lys). To form the precursor, a mass of 10 mg peptide preparation was dissolved in 800 μL sterile water (Sigma, W3500), using a 3 min vortex step followed by centrifugation (3 min at 1000 rpm) and a 2 h incubation at 80 °C. After incubation, 0.5 M NaOH (Sigma, S2770) was added incrementally to the gel until optically clear. The gel was vortexed, buffered by addition of 100 μL 10× PBS (Gibco, 70011), and incubated at 80 °C overnight. The resulting precursor could be stored at 4 °C until required.

Mendonca T., Lis-Slimak K., Matheson A.B., Smith M.G., Anane-Adjei A.B., Ashworth J.C., Cavanagh R., Paterson L., Dalgarno P.A., Alexander C., Tassieri M., Merry C.L, & Wright A.J. (2023). OptoRheo: Simultaneous in situ micro-mechanical sensing and imaging of live 3D biological systems. Communications Biology, 6, 463.

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Octapeptide Hydrogel Precursor Preparation

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The precursor and gel preparation method was followed as previously published 57 (link) . A commercially available peptide preparation in powder form was used as the source of the octapeptide gelator (Pepceuticals UK, FEFEFKFK, Phe-Glu-Phe-Glu-Phe-Lys-Phe-Lys). To form the precursor, a mass of 10 mg peptide preparation was dissolved in 800 μL sterile water (Sigma, W3500), using a 3 min vortex step followed by centrifugation (3 min at 1000 rpm) and a 2 hour incubation at 80 °C. After incubation, 0.5 M NaOH (Sigma, S2770) was added incrementally to the gel until optically clear. The gel was vortexed, buffered by addition of 100 μL 10x PBS (Gibco, 70011), and incubated at 80 °C overnight. The resulting precursor could be stored at 4 °C until required.

Mendonca T., Lis-Slimak K., Matheson A.B., Smith M.G., Anane‐Adjei A.B., Ashworth J., Cavanagh R., Paterson L., Dalgarno P.A., Alexander C., Tassieri M., Merry C.L., & Wright A.J. (2022). OptoRheo: Simultaneousin situmicro-mechanical sensing and imaging of live 3D biological systems.

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