Synergy 2 sl luminescence microplate reader

Manufactured by Agilent Technologies

Sourced in United States

The Synergy 2 SL Luminescence Microplate Reader is a laboratory equipment designed for the detection and measurement of luminescence signals. It is capable of reading multi-well microplates and provides accurate and sensitive luminescence detection.

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Lab products found in correlation

Rpmi 1640 without phenol red, by Merck Group (2 mentions) 2 7 dichlorofluorescein diacetate, by Thermo Fisher Scientific (2 mentions) Dihydrorhodamine 123, by Merck Group (1 mentions) Rpmi without phenol red, by Merck Group (1 mentions) 2 7 dichlorofluorescin diacetate dcfh da, by Thermo Fisher Scientific (1 mentions) Luciferase lysis buffer, by Promega (1 mentions) Luciferase reagent, by Promega (1 mentions) Transam nfκb p65 chemi kit, by Active Motif (1 mentions)

Close spelling variants of this product

Luminescence microplate reader Synergy™ 2 SL Synergy microplate reader Synergy 2 reader Luminescence reader Microplate reader Synergy 2

7 protocols using synergy 2 sl luminescence microplate reader

Quantifying Fungal ROS and Peroxynitrite

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The endogenous production of ROS and peroxynitrite by fungal cells was measured by fluorometry (Synergy 2 SL Luminescence Microplate Reader; BioTek^®) with specific probes (Ferreira et al., 2013 (link)). The cells (1.0 × 10³ to 5.0 × 10³ cells per milliliter) were incubated with EAF (MIC and 2 × MIC) or AMB (MIC) in RPMI 1640 without phenol red (Sigma-Aldrich^®) containing 10 mM 2’,7’-dichlorofluorescein diacetate (for ROS quantification; Invitrogen^®) or 20 mM dihydrorhodamine 123 (for peroxynitrite quantification; Sigma-Aldrich^®). The fluorescence was measured 24 h later at 500 nm. At the end of the experiments, 10 μl of each sample was placed on SDA-containing plates, and the numbers of CFU were counted. The results are expressed as arbitrary units of fluorescence/CFU.

Folly M.L., Ferreira G.F., Salvador M.R., Sathler A.A., da Silva G.F., Santos J.C., dos Santos J.R., Nunes Neto W.R., Rodrigues J.F., Fernandes E.S., da Silva L.C., de Freitas G.J., Denadai Â.M., Rodrigues I.V., Mendonça L.M., Monteiro A.S., Santos D.A., Cabrera G.M., Siless G, & Lang K.L. (2020). Evaluation of in vitro Antifungal Activity of Xylosma prockia (Turcz.) Turcz. (Salicaceae) Leaves Against Cryptococcus spp.. Frontiers in Microbiology, 10, 3114.

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Measuring Endogenous ROS in Candida

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Endogenous ROS measured by fluorometric assay with specific probes (Ferreira et al., 2013 (link)). Yeast cells from C. gattii and C. neoformans cultured on SDA (37°C for 72 h) were treated with (MIC eugenol = 256 mg/L for C. gattii ATCC 24065, C. neoformans ATCC 28957, and C. neoformans ATCC 62066; 64 mg/L – C. gattii ATCC 32068) and hydrogen peroxide (positive control) for 1 h in RPMI without phenol red (Sigma–Aldrich, St Louis, MO, United States) and incubated with 10 μM of 2′,7′-dichlorofluorescin diacetate (DCFH-DA; Invitrogen, Life Technologies, Carlsbad, CA, United States) for ROS quantification. A growth control was also performed. The fluorescence was measured with a Fluorometer (Synergy 2 SL Luminescence Microplate Reader; Biotek) using excitation and emission wavelengths of 500 nm. The results were expressed as arbitrary units of fluorescence ±SEM. These tests were performed in triplicate.

Alves J.C., Ferreira G.F., Santos J.R., Silva L.C., Rodrigues J.F., Neto W.R., Farah E.I., Santos Á.R., Mendes B.S., Sousa L.V., Monteiro A.S., dos Santos V.L., Santos D.A., Perez A.C., Romero T.R., Denadai Â.M, & Guzzo L.S. (2017). Eugenol Induces Phenotypic Alterations and Increases the Oxidative Burst in Cryptococcus. Frontiers in Microbiology, 8, 2419.

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Quantifying HIV-1 Infectivity in TZM-bl Cells

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Utilizing the TZM-bl cell line, a HeLa cell derivative that are CD4+ and expresses CCR5 and CXCR4, as well as a Tat-inducible luciferase reporter, HIV-1 infectivity was quantified on the basis of relative luciferase expression.^{12 (link),18 (link)} These cells, which are permissive to wild type HIV-1, were pre-exposed to PHT-conditioned media or non-conditioned media for 24 h prior to infection. HIV-1 NL4-3 was added at increasing concentrations, indicated in the figure legend. After 48 h, infected cells were washed with PBS and lysed in luciferase lysis buffer (Promega, USA). Lysates (40 μl) were transferred to white 96-well plates, and 50 μl luciferase reagent (Promega) was injected into each well. Luciferase activity was determined by detection of luminescence recorded by Synergy 2 SL luminescence microplate reader (BioTek, VT, USA).

BAYER A., DELORME-AXFORD E., SLEIGHER C., FREY T.K., TROBAUGH D.W., KLIMSTRA W.B., EMERT-SEDLAK L.A., SMITHGALL T.E., KINCHINGTON P.R., VADIA S., SEVEAU S., Boyle J.P., COYNE C.B, & SADOVSKY Y. (2014). Human trophoblasts confer resistance to viruses implicated in perinatal infection. American journal of obstetrics and gynecology, 212(1), 71.e1-71.e8.

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NF-κB p65 Activation Assay in CLL

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The TransAM^® NF-κB p65 Chemi kit (Active Motif, Carlsbad, CA) was used to detect NF-κB activity. TG02 or Lcki were added 1 h prior to anti-IgM, and the CLL cells were collected 2 h later. Two microgram nuclear extracts were used for p65 activation analysis by ELISA per kit instruction. Nuclear extract from TPA and calcium ionophore activated Jurkat cells were used as positive control. The wild-type and mutated p65 consensus binding oligonucleotides were used to monitor the specificity of the analysis. The chemiluminescence was read with a Synergy2 SL Luminescence Microplate Reader (BioTek, Winooski, VT).

Chen R., Tsai J., Thompson P.A., Chen Y., Xiong P., Liu C., Burrows F., Sivina M., Burger J.A., Keating M.J., Wierda W.G, & Plunkett W. (2021). The multi-kinase inhibitor TG02 induces apoptosis and blocks B-cell receptor signaling in chronic lymphocytic leukemia through dual mechanisms of action. Blood Cancer Journal, 11(3), 57.

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Quantifying Protein Immobilization Kinetics

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Bovine serum albumin (BSA, Invitrogen) conjugated with Alexa Fluor 488 was chosen as a model protein to study the dissociation kinetics of immobilized proteins. A 0.001% (w/v) solution was prepared and briefly centrifuged to remove any protein aggregates, which can lead to nonspecific background fluorescence. Fiber samples on 1 cm² silicon chips were placed in a 12-well plate, incubated with 20 μg of BSA for 90 min, and then washed with 1× PBS three times. Fresh 1× PBS was added to each well before the relative intensity was read with the Synergy 2 SL Luminescence Microplate Reader (BioTek, VT). Samples were stored in the dark at room temperature between reads. The ability of each sample to sequester the proteins was evaluated for six days. The observed relative intensities of the immobilized BSA were converted to concentrations with the use of equations derived from a known concentration ladder.

Cohn C., Leung S.L., Zha Z., Gamboa J., Teng W, & Wu X. (2015). Comparative Study of Antibody Immobilization Mediated by Lipid and Polymer Fibers. Colloids and surfaces. B, Biointerfaces, 134, 1-7.

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Antibody Immobilization on Electrospun Fibers

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The immobilization of anti-CD20 antibodies on the CSS-PCL fibers was examined, in comparison to the PCL and plasma-treated PCL controls. Following a protocol previously reported [19 (link), 26 (link)], silicon chips that collected electrospun fibers were each placed in a single well of a 48-well plate, rinsed with PBS three times, exposed to 200 μL of a 10 μg/mL anti-CD20 solution, and incubated for 90 min at 37 °C. After the incubation period the solution was collected and placed in fresh wells of a new and sterile 96-well plate. The fiber specimens and wells were rinsed three times with PBS. The amount of unbound anti-CD20 remaining in the solution and washes from the same specimen was detected with a microplate reader (Synergy 2 SL Luminescence Microplate Reader, BioTek, Winooski, USA) at an excitation wavelength of 460/40 nm and an emission wavelength of 590/10 nm.

Cohn C., Leung S.L., Crosby J., Lafuente B., Zha Z., Teng W., Downs R, & Wu X. (2016). Lipid-mediated protein functionalization of electrospun polycaprolactone fibers. EXPRESS polymer letters, 10(5), 430-437.

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Quantification of Macrophage ROS

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BMDMs were cultured in RPMI1640 without phenol red (Sigma-Aldrich), treated or not with 10-nM E2, infected with Cg, and incubated with 10-μM 2′,7′-dichlorofluorescein diacetate (Invitrogen, Life Technologies, Carlsbad, CA, USA) for ROS quantification. Two and 24 hr post inoculation, fluorescence was assessed by using a fluorometer (Synergy 2 SL Luminescence Microplate Reader; Biotek) using excitation and emission wavelengths of 500 nm. Data are expressed as the mean of arbitrary units of fluorescence of six technical replicates (Ferreira et al., 2013) and are representative of three independent experiments.

Costa M.C., de Barros Fernandes H., Gonçalves G.K.N., Santos A.P.N., Ferreira G.F., de Freitas G.J.C., do Carmo P.H.F., Hubner J., Emídio E.C.P., Santos J.R.A., Dos Santos J.L., Dos Reis A.M., Fagundes C.T., da Silva A.M, & Santos D.A. (2020). 17-β-Estradiol increases macrophage activity through activation of the G-protein-coupled estrogen receptor and improves the response of female mice to Cryptococcus gattii. Cellular microbiology, 22(6).

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