Synergy ht multi detection microplate reader

Manufactured by Agilent Technologies

Sourced in United States, Germany, Switzerland

The Synergy HT Multi-Detection Microplate Reader is a versatile laboratory instrument designed for high-throughput microplate-based assays. It is capable of performing absorbance, fluorescence, and luminescence detection.

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Lab products found in correlation

Bca protein assay, by Thermo Fisher Scientific (8 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (7 mentions) Z phe arg amc, by Merck Group (4 mentions) Sytox green, by Thermo Fisher Scientific (4 mentions) Atplite luminescence assay system, by PerkinElmer (3 mentions) Folin ciocalteu reagent, by Merck Group (3 mentions) Gen5tm, by Agilent Technologies (3 mentions) Trizol, by Thermo Fisher Scientific (3 mentions) Mtt assay, by Merck Group (2 mentions) Magpix multiplex fluorescence reader, by DiaSorin (2 mentions) Bio plex manager mp software, by DiaSorin (2 mentions) Brilliant 2 sybr green qpcr master mix kit, by Agilent Technologies (2 mentions) Aria mix real time pcr system, by Agilent Technologies (2 mentions) Ariamx 1.0 program, by Agilent Technologies (2 mentions) Affinityscript qpcr cdna synthesis kit, by Agilent Technologies (2 mentions) Take3 multi volume plate, by Agilent Technologies (2 mentions) Alcian blue, by Merck Group (2 mentions) Celltiter glo reagent, by Promega (2 mentions) Caspase glo 9 assay kit, by Promega (2 mentions) Caspase glo 3 7, by Promega (2 mentions)

243 protocols using synergy ht multi detection microplate reader

Evaluating Cell Viability and Apoptosis

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The cell viability of RAW264.7 cells was determined by MTT assay. RAW264.7 cells were seeded on 96-well culture plates at density of 1 × 10⁵cells/ml and pretreated with various concentrations of LREE. And the cells were added with 20 μL of MTT (5 mg/ml) each well and incubated for additional 4 h. Subsequently, the supernatant was removed and the formazone crystals were dissolved using DMSO 150 μL. The light absorbance at 540 nm was quantified with a Bio-Tek Synergy HT Multi-Detection Microplate Reader.
The cell viability of CD4⁺ T cells stimulated with anti-CD3 and anti-CD28 was determined by Cell Counting Kit 8 assay (CCK-8). The CD4⁺ T cells were seeded on 96-well culture plates at density of 5 × 10⁵ cells/ml and pretreated with various concentrations of LREE. And the cells were added with 10 μL of CCK-8 regent each well and incubated for additional 4 h. The light absorbance at 450 nm was quantified with a Bio-Tek Synergy HT Multi-Detection Microplate Reader.
The cells were transferred into the flow tube, washed with PBS, then centrifuged and discarded the supernatant. After resuscitating the cells with PBS, the cells were incubated for 5 min at room temperature in the dark with 5 μL Annexin V-APC and 10 μL PI. Finally, the samples were detected by flow cytometry.

Lai H., Yang Z., Lou Z., Li F., Xie F., Pan W., Xu C., Zhang L., Zhang S., Zhang L, & Huang M. (2021). Root Extract of Lindera aggregata (Sims) Kosterm. Modulates the Th17/Treg Balance to Attenuate DSS-Induced Colitis in Mice by IL-6/STAT3 Signaling Pathway. Frontiers in Pharmacology, 12, 615506.

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Antioxidant Activity Evaluation of TEE

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To estimate the antioxidant activity of TEE, DPPH radical and hydrogen peroxide scavenging activities were determined according to previously reported methods [40 (link),41 (link)]. Briefly, a solution of 4 × 10⁴ M DPPH in methanol was prepared, and 100 µL of this solution was mixed with 100 µL of TEE in multiple concentrations. The mixtures were incubated in a shaking incubator for 30 min at 25–30 °C. After incubation, the absorbance was measured spectrophotometrically at 517 nm (Synergy HT Multi-Detection microplate reader, BioTek, Winooski, VT, USA). For hydrogen peroxide scavenging activity, 100 µL of TEE in multiple concentrations and 0.1 M phosphate buffer (pH 5.0) was mixed with 10 mM hydrogen peroxide solution and incubated for 5 min at 37 °C. Then, 30 μL of 1.25 mM ABTS and 1 unit/mL peroxidase were added. The mixtures were incubated for 10 min at 37 °C, and the incubated product was measured at 405 nm using a microplate reader (Synergy HT Multi-Detection microplate reader, BioTek, Shoreline, DC, USA).

Kim E.A., Kang N., Heo S.Y., Oh J.Y., Lee S.H., Cha S.H., Kim W.K, & Heo S.J. (2023). Antioxidant, Antiviral, and Anti-Inflammatory Activities of Lutein-Enriched Extract of Tetraselmis Species. Marine Drugs, 21(7), 369.

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Evaluating Fucoxanthin-Treated PL-MSC Viability

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The viability of fucoxanthin-treated PL-MSCs was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Sigma-Aldrich, USA). Briefly, PL-MSCs were seeded at a density of 3 × 10⁴ cells/cm² into a 96-well plate (Corning, USA). After overnight culture, cells were treated with fucoxanthin (Sigma-Aldrich, USA) at final concentrations of 1, 2, 3, 4, 5, 10, and 20 µM for 24, 48 and 72 h. Following the respective experimental time, the cells were incubated with 0.5 mg/ml MTT in completed DMEM for 4 h at 37 °C. Subsequently, 100 μl dimethyl sulfoxide (DMSO) was added to each well. The absorbance was measured at 570 nm using a Synergy HT Multi-Detection Microplate Reader and Agilent BioTek Gen6 software (BioTek Instruments Inc., USA). Cell viability was calculated as shown in the equation:

Cell viability (% of control) = (\frac{OD (test) - OD (blank)}{OD (control) - OD (blank)}) \times 100

Suwanmanee G., Tantrawatpan C., Kheolamai P., Paraoan L, & Manochantr S. (2023). Fucoxanthin diminishes oxidative stress damage in human placenta-derived mesenchymal stem cells through the PI3K/Akt/Nrf-2 pathway. Scientific Reports, 13, 22974.

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Exploring G. murorum Laccase Thermostability

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To test the dependence of G. murorum laccase activity on temperature, enzymatic assays were performed in a 96‐well flat‐wells plate (ThermoFisher Scientific) whose wells were filled with 398 μl of laccase reaction buffer (100 mM sodium acetate buffer, pH 4.5; 0.2 mM ABTS) used in triplicate for the test (enzymatic assays) and control (buffer along) samples. Once the plate was preheated in the plate‐reader (Synergy™ HT Multi‐Detection Microplate Reader; BioTek®; Agilent) to 20°C, 30°C, 40°C or 50°C, 2 μl of G. murorum protein sample (prepared as samples used for zymography) was added to the test samples and the absorbance of all samples was read for 20 min with 1 min intervals.

Fernández‐Remacha D., González‐Riancho C., Lastra Osua M., González Arce A., Montánchez I., García‐Lobo J.M., Estrada‐Tejedor R, & Kaberdin V.R. (2022). Analysis of laccase‐like enzymes secreted by fungi isolated from a cave in northern Spain. MicrobiologyOpen, 11(2), e1279.

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Determination of Total Phenolic Content in Blueberry Pomace

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Total phenolic content of the blueberry pomace extracts was determined using the method of Folin–Ciocalteu following the procedure described by Tournour et al. [42 (link)] with slight modification. Briefly, 25 μL of either sample or standard properly diluted with milliQ water were transferred into appropriate wells. With a multichannel pipet, 125 μL of 0.2 N Folin–Ciocalteu’s reagent were added to each well, then the plate was swirled and incubated in the dark at room temperature. After 8 to 10 min, 125 μL of 7.5% sodium carbonate was added. The obtained solution was mixed thoroughly and incubated at room temperature for 30 min at least and no more than 60 min. Subsequently, the absorbance was recorded at 765 nm with a spectrophotometric microplate reader (Synergy HT Multi-Detection Microplate Reader, BioTek Instruments, Winooski, VT, USA). Absorbance was compared to a gallic acid standard curve (R² = 0.999) to quantify TPC in the sample. The results were expressed as milligrams of gallic acid equivalents per gram of dry matter (mg GAE/g DM). Each standard and sample solution was analysed in triplicate.

Bamba B.S., Shi J., Tranchant C.C., Xue S.J., Forney C.F, & Lim L.T. (2018). Influence of Extraction Conditions on Ultrasound-Assisted Recovery of Bioactive Phenolics from Blueberry Pomace and Their Antioxidant Activity. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(7), 1685.

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Endothelial Biomarkers Quantification

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Circulating levels of E-selectin, P-selectin, and the enzyme endothelial nitric oxide synthase (eNOS) were determined in plasma by ELISA following the protocols of Cloud-Clone Corp. (Katy, TX, USA). Spectrophotometric reading was performed using a Bio-Tek^® Synergy™ HT Multi-Detection microplate reader controlled by BioTek^®Gen5 software version 2.01.14 (BioTek Instruments, Winooski, VT, USA). Intercellular (ICAM-1) and vascular (VCAM-1) adhesion molecules were determined in serum samples with Bio-Plex^® Pro Human Cytokine ICAM-1 and VCAM-1 kits (Bio-Rad, Hercules, CA, USA) using a MAGPIX™ Multiplex fluorescence reader operating with the Bio-Plex Pro Wash Station and the Bio-Plex Manager™ MP software for data processing (Luminex Corporation, Austin, TX, USA).

González-Rámila S., Sarriá B., Seguido M.Á., García-Cordero J., Bravo-Clemente L, & Mateos R. (2022). Effect of Olive Pomace Oil on Cardiovascular Health and Associated Pathologies. Nutrients, 14(19), 3927.

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Spectrophotometric Quantification of Urinary Creatinine

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The creatinine concentration of each urinary sample was spectrophotometrically quantified using the QuantiChrom™ Creatinine Assay Kit (BioAssay Systems). In this assay, creatinine reacts with picric acid in an alkaline solution forming a yellow orange adduct. The increase in absorbance of this adduct is measured at a wavelength of 510 nm. For the assay, 10 μl of mouse urine was mixed with 200 μl working solution (0.1 M sodium hydroxide (NaOH); 0.3 mM ethylenediaminetetraacetic acid (EDTA); 0.1% (v/v) DMSO; 0.0004% (v/v) Tween®20; and 3 mM picric acid), in duplicate, in a 96-well plate. The samples were then analysed via a Synergy™ HT Multi-detection microplate reader (Biotek® Instruments) by assessing the increase in linear trend of absorbance at 510 nm for 5 min, in 1 min intervals. Absolute quantities were determined by making use of a standard curve, ranging from 0 to 50 mg%.

Willemse L., Terburgh K, & Louw R. (2023). A ketogenic diet alters mTOR activity, systemic metabolism and potentially prevents collagen degradation associated with chronic alcohol consumption in mice. Metabolomics, 19(5), 43.

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Cell Viability Measurement by ATPlite Assay

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Cell viability was measured by the ATPlite Luminescence Assay System (PerkinElmer, Waltham, MA, USA), as per manufacturer’s instructions, using a Synergy HT Multi-Detection Microplate Reader (Bio-Tek Instruments, Winooski, VT, USA) to measure the relative luminescence units (RLU). The RLUs of untreated cells was considered as 100% viability; the results were expressed as a percentage of viable cells versus untreated cells.

Belisario D.C., Akman M., Godel M., Campani V., Patrizio M.P., Scotti L., Hattinger C.M., De Rosa G., Donadelli M., Serra M., Kopecka J, & Riganti C. (2020). ABCA1/ABCB1 Ratio Determines Chemo- and Immune-Sensitivity in Human Osteosarcoma. Cells, 9(3), 647.

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Quantifying Polyphenols in Cyanobacterial Extracts

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The TPC of the cyanobacterial extracts was determined using the colorimetric assay of Folin–Ciocalteu, according to Barroso et al. [18 (link),48 (link)]. The acetonic extracts were solubilized in DMSO and the aqueous extracts in water. Briefly, a volume of 25 μL of each extract (10 mg mL⁻¹) was thoroughly mixed with 25 μL of Folin–Ciocalteu reagent (Sigma-Aldrich, St. Louis, MO, USA), 200 μL of Na₂CO₃ solution (75 g L⁻¹) and 500 μL of deionized water. After the incubation period (60 min at room temperature), the absorbance of the colored product was measured at 725 nm, using a Synergy HT Multi-detection microplate reader (Biotek, Bad Friedrichshall, Germany) operated by GEN5^TM software. Standard calibration curves (y = 2.097x + 0.01560, R² = 0.9989, for aqueous extracts and y = 2.204x + 0.01401, R² = 0.9982, for acetonic extracts) were obtained with seven concentrations of gallic acid (GA) (0.025 to 0.5 mg mL⁻¹). TPC in each extract was expressed in µg of gallic acid equivalents (GAE) per mg of dry biomass. Three independent determinations were carried out in triplicate.

Morone J., Lopes G., Morais J., Neves J., Vasconcelos V, & Martins R. (2022). Cosmetic Application of Cyanobacteria Extracts with a Sustainable Vision to Skincare: Role in the Antioxidant and Antiaging Process. Marine Drugs, 20(12), 761.

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Quantitative RT-PCR Analysis of Epithelial-Mesenchymal Transition Genes

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Cells were grown to confluence and lysed by adding 1 ml of TRIZOL (Thermo Fisher, Waltham, MA, USA; Cat. 15596-026) directly in the culture dish (1 ml per 3.5 cm diameter dish) and scraping. RNA was extracted and quantified using a Synergy HT Multi-Detection Microplate Reader (BIOTEK, Winooski, VT, USA). cDNA was synthesized using the AFFINITYSCRIPT QPCR CDNA synthesis kit (Agilent Tech., Santa Clara, CA, USA; Cat. 600559) following the manufacturer's protocol. qRT-PCR was performed in triplicate using the Brilliant II SYBR green QPCR Master Mix kit (Agilent Tech., Cat. 600828) in an Aria Mix Real-Time PCR system (Agilent Tech., 68830A model). Data were analyzed using the AriaMX 1.0 program (Agilent Tech.). The following sets of primers were used: ZEB1 (s): 5'-GTA AGA GGC CTC ACG AGT GT-3', (as): 5'-GCA GTA GGA GTA GCG GTG AT -3', CDH1 (E-Cadherin) (s): 5'-GAA CGC ATT GCC ACA TAC AC-3', (as): 5'-ATT CGG GCT TGT TGT CAT TC-3', KRT18 (Cytokeratin 18) (s): 5'-ACA GAG TGA GGA GCC TGG AGA CCG A-3', (as): 5'-CAG TAT TTG CGA AGA TCT GAG CCC TC-3', Cdh2 (N-Cadherin) (s): 5'-GGA CAG TTC CTG AGG GAT CA-3', (as): 5'-GGA TTG CCT TCC ATG TCT GT-3', VIM (Vimentin) (s): 5'-GCC AAG GCA AGT CGC G-3', (as): 5'- CA TTT CAC GCA TCT GGC G-3'. The mRNA levels were normalized using the housekeeping gene PUM1 (Pumilio) (s): 5'-CGG TCG TCC TGA GGA TAA A-3', (as): 5'-CGT ACG TGA GGC GTG AGT AA-3'.

Orellana-Serradell O., Herrera D., Castellon E.A, & Contreras H.R. (2017). The transcription factor ZEB1 promotes an aggressive phenotype in prostate cancer cell lines. Asian Journal of Andrology, 20(3), 294-299.

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