Synergy 2 multi mode plate reader

Manufactured by Agilent Technologies

Sourced in United States

The Synergy 2 Multi-Mode Plate Reader is a versatile laboratory instrument designed for conducting various assays and measurements. It is capable of detecting and quantifying a wide range of analytes, including cells, proteins, nucleic acids, and small molecules, across multiple detection modes such as absorbance, fluorescence, and luminescence.

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Synergy 2 Multi-Mode BioTek plate reader Synergy Multi-Mode Plate Reader Synergy 2 Multi-Mode Reader Synergy 2 Multi-Mode Synergy 2 plate reader Synergy plate reader Synergy 2 reader Synergy 2 plate Synergy 2

59 protocols using synergy 2 multi mode plate reader

Rigosertib Sensitivity in PDX Organoids

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LU-NB-1, LU-NB-2, and LU-NB-3 PDX-derived tumor organoids were dissociated into single cells and seeded into opaque 96-well plates (Corning Inc., Corning, NY), 5000 cells per well, and treated immediately with a range (0–100 nM) of rigosertib concentrations. Cells were incubated for 72 h. Cell viability was calculated as a percentage of control wells based on CellTiter-Glo (G7571; Promega, Madison, WI) luminescence. Luminescence was measured with a Synergy2 Multi-Mode plate reader (BioTek, Winooski, VT). Biological triplicates were used.

Radke K., Hansson K., Sjölund J., Wolska M., Karlsson J., Esfandyari J., Pietras K., Aaltonen K., Gisselsson D, & Bexell D. (2021). Anti-tumor effects of rigosertib in high-risk neuroblastoma. Translational Oncology, 14(8), 101149.

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Fluorescent Polarization Assay for Protein Binding

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The C8 substrate sense-strand oligo was synthesized with a 5′ Alexa-488 fluorescent dye (5′-Alex488N-GACCATGCTTAGGGTTAGGGTTATCATACAAGTTAGGGTTAGGG-3′) and thermally annealed with the anti-sense strand (5′-TTGTATGATAACCCTAACCCTAAGCATGGTC-3′) (Integrated DNA Technologies). Each 40 µl reaction (buffer—50 mM Tris-HCl pH 8.0, 150 mM KCl, 2 mM MgCl₂, 1 mM DTT, 10% (v/v) glycerol, 0.5 mg ml⁻¹ BSA) contained 5 nM (500 nM for saturation-binding assay) fluorescent substrate and was incubated with protein for 30 min at 23 °C before fluorescent polarization measurement in a 384-well plate with Synergy 2 multi-mode plate reader (BioTek). For competitor assays, initial measurements were taken (defined as t = 0) before adding competitors and taking measurements every 2 min for 4 h.

Lim C.J., Zaug A.J., Kim H.J, & Cech T.R. (2017). Reconstitution of human shelterin complexes reveals unexpected stoichiometry and dual pathways to enhance telomerase processivity. Nature Communications, 8, 1075.

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Caspase Inhibition in Smooth Muscle Cells

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To test the efficacy of Z-VAD(OH)-FMK, a pan-caspase inhibitor with specific affinity for the final apoptotic effector caspase 6, rat smooth muscle primary cells (SMAC) were cultured in sterile 96-well dark plates until confluent and incubated for 30 min at 37 °C and subsequently cooled to 4 °C for 30, 60, 90, 120, 150, and 180 min. Caspase activity was assessed using the Caspase-Glo^® 3/7 Assay (Promega, Madison, WI, USA) according to manufacturer’s protocol, and luminescence was measured using a Synergy 2 Multi-Mode plate reader (BioTek). Data were expressed as total luminescence absorbed. A two-way ANOVA with Tukey’s multiple comparisons test was used for statistical analysis.

Gartzke L.P., Hendriks K.D., Hoogstra-Berends F., Joschko C.P., Strandmoe A.L., Vogelaar P.C., Krenning G, & Henning R.H. (2023). Inhibition of Ferroptosis Enables Safe Rewarming of HEK293 Cells following Cooling in University of Wisconsin Cold Storage Solution. International Journal of Molecular Sciences, 24(13), 10939.

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Geopolymer Pots for Wheat Growth

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Seeds of spring wheat, var. Almari, were germinated and grown for ten days in pots filled with small pieces of cellulose pats. The plant’s morphology was compared for plants grown in geopolymer pots vs plastic pots. Ion adsorption and desorption properties of the geopolymer foam pots were tested. The pots were immersed for 24 h in the mineral solution described above. Ten subsequent rinses with distilled water were performed. At each step, pots were filled with 50 mL distilled water and after 24 h, the leaches were analysed spectrophotometrically (Synergy 2 Multi-Mode Plate Reader, BioTek Instruments, Winooski, VT, USA). The procedures were carried out in accordance with the manufacturer’s instructions using commercial kits ZW 535550 (Slandi Ltd. Warsaw, Poland), 1-403-B125, 1-440-0080 (BioMaxima S.A., Lublin, Poland) for nitrogen, phosphorus, and potassium analysis, respectively, and standards for their content quantification (Química Clínica Aplicada S.A.).

Szechyńska-Hebda M., Marczyk J., Ziejewska C., Hordyńska N., Mikuła J, & Hebda M. (2019). Optimal Design of pH-neutral Geopolymer Foams for Their Use in Ecological Plant Cultivation Systems. Materials, 12(18), 2999.

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Protein Extraction from Tissue Samples

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LC and DR tissue punches were homogenized in 200 μL of tissue lysis buffer (500 mM NaCl, 50 mM Tris HCl pH 7.6, 10% NP-40, 70% glycerol) with 0.02% protease phosphatase inhibitor cocktail (Thermo Scientific, Waltham, MA) in conjunction with mechanical disruption with a bullet blender (Next Advance, Averill Park, NY) at 4°C. Protein concentrations of the supernatant were assessed using a BCA assay (Thermo Scientific, Waltham, MA) according to the manufacturer’s protocol and read using a Synergy 2 Multi-Mode plate reader (Bio Tek, Winooski, VT) with Gen5 software (Bio Tek, Winooski, VT). The remaining tissue homogenate was stored at −80°C.

Finnell J.E., Lombard C.M., Melson M.N., Singh N.P., Nagarkatti M., Nagarkatti P., Fadel J.R., Wood C.S, & Wood S.K. (2016). The protective effects of resveratrol on social stress-induced cytokine release and depressive-like behavior. Brain, behavior, and immunity, 59, 147-157.

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Fluorescence Polarization Assay for PRC2

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Each dsDNA containing 5’ Alexa-488 fluorescent dye was synthesized, annealed, and purified by IDT. Each 40 µl reaction contained 5 nM fluorescent DNA and PRC2 in binding buffer (50 mM Tris-HCl pH 7.5, 25 mM KCl, 2.5 mM MgCl₂, 0.1 mM ZnCl₂, 2 mM 2-mercaptoethanol, 0.1 mg/ml bovine serum albumin, 5% v/v glycerol). Reaction was incubated for 30 min at 30°C, followed by measuring fluorescence polarization in a 384-well plate with Synergy 2 multi-mode plate reader (BioTek). For competitor assays, initial signals were measured (defined as t = 0) before adding competitors. RNA or dsDNA cold competitors were added into the reactions and the measurements were taken every 20 s for 4 h.

Wang X., Paucek R.D., Gooding A.R., Brown Z.Z., Ge E.J., Muir T.W, & Cech T.R. (2017). Molecular analysis of PRC2 recruitment to DNA in chromatin and its inhibition by RNA. Nature structural & molecular biology, 24(12), 1028-1038.

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Oxalic acid extraction of α-elastin

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Cell cultures were lysed with RIPA buffer (ThermoFisher), and cellular and extracellular material was collected using cell scrapers (Biologix), sonicated, and centrifuged to separate soluble and insoluble material. Pelleted material was then subjected to two 1 hour hot 0.25 M oxalic acid treatments at 100°C to extract digested α-elastin soluble derivatives. Samples were then analyzed via the Fastin^™ assay (Biocolor), following the manufacturer’s instructions, with absorbances measured at 513 nm using a Synergy 2 multi-mode plate reader (BioTek).

Ellis M.W., Riaz M., Huang Y., Anderson C.W., Luo J., Park J., Lopez C.A., Batty L.D., Gibson K.H, & Qyang Y. (2021). Epigallocatechin Gallate Facilitates Extracellular Elastin Fiber Formation in Induced Pluripotent Stem Cell Derived Vascular Smooth Muscle Cells for Tissue Engineering. Journal of molecular and cellular cardiology, 163, 167-174.

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Quantifying HCV Replication and Infectivity

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5×10⁶ cells were electroporated with 10 μg HCV genomic RNA as described previously (56 (link)) and plated to 25 cm² cell culture vessels for assays to assess intracellular viral RNA abundance (GLuc assay) or extracellular infectious virus production (FFU assay). For GLuc assays, cell culture medium was harvested and replaced at 6, 24, 48 and 72 hours after electroporation. Cell culture medium was analyzed for GLuc activity using the BioLux Gaussia substrate (New England BioLabs) on a Synergy 2 Multimode Plate Reader (Biotek). For FFU assay, cell culture medium was harvested and replaced at 24, 48 and 72 hours after electroporation. Infectious virus was measured as described previously (56 (link)).

Doncheva N.T., Domingues F.S., McGivern D.R., Shimakami T., Zeuzem S., Lengauer T., Lange C.M., Albrecht M, & Welsch C. (2019). Near neighbor interactions in the NS3–4A protease of HCV impact replicative fitness of drug-resistant viral variants. Journal of molecular biology, 431(12), 2354-2368.

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hiPSC-VSMC Proliferation Assay

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hiPSC-VSMCs derived from EB-based or chemically defined approach were seeded at 20,000 cells/well density into GFR-Matrigel-coated 96-well plates with expansion medium and cultured for three days. Cell proliferation was measured as a function of metabolic activity using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) (Sigma-Aldrich) on Day 4. MTT at 0.5 mg/ml was added into the medium of each well and incubated with the cells at 37°C for 2 hours, follo wed by cell solubilization with DMSO (AmericanBIO) for 15 minutes. Absorbance was measured at 540 nm using the Synergy 2 multi-mode plate reader (BioTek). Three biological replicates were completed for evaluation of each cell group.

Luo J., Qin L., Zhao L., Gui L., Ellis M.W., Huang Y., Kural M.H., Clark J.A., Ono S., Wang J., Yuan Y., Zhang S.M., Cong X., Li G., Riaz M., Lopez C., Hotta A., Campbell S., Tellides G., Dardik A., Niklason L.E, & Qyang Y. (2020). Tissue Engineered Vascular Grafts with Advanced Mechanical Strength from Human iPSCs. Cell stem cell, 26(2), 251-261.e8.

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Glutathione Quantification in THP-1 Cells

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Reduced glutathione (GSH), oxidized glutathione (GSSG), and total glutathione levels were determined in THP-1 cells following treatment. Glutathione was determined based on modifications of the Tietze recycling assay [22] (link). In brief, cells were lysed in lysis buffer containing 0.1% Triton X-100 in PBS buffer, pH 7.4, containing 10 µM DTPA. Cell lysates were treated with triethanolamine (TEA), 2-methyl-5-vinylpyridine (MVP), and 5% 5-sulfosalicylic acid (5-SA) to measure GSSG or 5-SA alone to measure GSH based on adapted methods from Anderson and Neufer [23] (link), [24] (link). Glutathione was determined based on the reduction of DTNB (Ellman's Reagent) at 412 nm in the Diabetes Research Center BioAnalytical Redox Biology Core (DK 079626) using a Synergy-2 Multimode plate reader (Biotek, Winooski, VT). Samples were normalized to cellular protein.

Patel M., Yarlagadda V., Adedoyin O., Saini V., Assimos D.G., Holmes R.P, & Mitchell T. (2017). Oxalate induces mitochondrial dysfunction and disrupts redox homeostasis in a human monocyte derived cell line. Redox Biology, 15, 207-215.

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