Synergy h4 microplate reader

Manufactured by Agilent Technologies

Sourced in United States

The Synergy H4 microplate reader is a versatile lab equipment designed for high-performance detection of a wide range of assays. It offers multi-mode detection capabilities, including absorbance, fluorescence, and luminescence, enabling researchers to conduct various biological and biochemical analyses.

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Synergy H4 Microplate Reader system (6 citations) Synergy H4 Hybrid Multi-Mode Microplate Reader (158 citations) Synergy H4 Hybrid Microplate Reader (130 citations) BioTek Synergy H4 Hybrid Multi-Mode Microplate Reader (5 citations) Synergy H4 Multi-Mode Microplate Reader (24 citations) Synergy™ H4 fluorescence microplate reader (2 citations) Bio Tek Synergy H4 Hybrid Microplate Reader (4 citations) Synergy H4 reader (18 citations) Synergy microplate reader (229 citations) Synergy H4 (609 citations)

92 protocols using synergy h4 microplate reader

Evaluating Compound Effects on TF-1 Leukemia Cells

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TF‐1/IDH2‐R140Q and TF‐1/IDH2‐R172K cells were treated with compounds for 7 days in medium with 2 ng/mL hGM‐CSF, and another 7 days without hGM‐CSF. At the end of treatment, MTT solution was added to each well, and plates were incubated for 4 hours at 37°C. Formazan crystals formed were then solubilized in acidic isopropanol, and the optical density of the resulting solution was read at 570 nm using a Synergy H4 microplate reader (BioTek).

Gao M., Zhu H., Fu L., Li Y., Bao X., Fu H., Quan H., Wang L, & Lou L. (2019). Pharmacological characterization of TQ05310, a potent inhibitor of isocitrate dehydrogenase 2 R140Q and R172K mutants. Cancer Science, 110(10), 3306-3314.

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Evaluating Cell Viability with Alkaloids

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To ascertain cell viability in
the presence of the isolated alkaloids, the assessment was conducted
using the CCK-8 kit.^{16 (link)} A 96-well microplate
was utilized to culture 3T3-L1 preadipocytes (1 × 10³ cells per well). Following a 12 h-incubation period, the cells were
exposed to varying concentrations of alkaloids for 48 h. The experimentation
was carried out following the specified guidelines provided by the
manufacturer. Subsequently, the optical density of the cell cultures
at 450 nm was quantified using a Synergy H4 microplate reader, which
was manufactured by BioTek (Winooski, VT).

Zhang P., Li J., Shi J., Cheng Z, & Wu T. (2024). Structurally Diverse Bisbenzylisoquinoline Alkaloids with Antiadipogenic Activity through PPARγ Downregulation from the Embryo of Nelumbo nucifera Seeds. Journal of Natural Products, 87(4), 1013-1022.

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Photosensitizer-Induced Cell Viability Assay

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5 × 10³ cells/well were seeded in complete medium on 96 well-plates 24 h before compounds treatment. BTI, T1, T2, and T3 were dissolved in complete medium at the indicated concentrations (DMSO reached the max value = 0.5 % v/v) and added to cells for 24 h at 37 °C in 5% CO₂. When required, the cells were photo-irradiated by using a EVOS® FL cell imaging system equipped with an adjustable-intensity LED cube (Blue light: excitation = 470/22 nm) operating at 23 mW cm⁻² for 6 min. T2- or T3-treated HeLa cells were also subjected to green or orange light irradiation, respectively. Adjustable-intensity LED cubes (Green light: excitation = 542/20 nm or Orange light: excitation = 585/29 nm) operating at 10 and 15.5 mW cm⁻² for 6 or 14 min, respectively, were used to deliver the light. Then, the cells were incubated for additional 24 h at 37 °C in 5% CO₂. At 48 h after compounds treatment, PrestoBlue (Invitrogen, Ref No: A13261) was added to each well and the cells were incubated at 37 °C in 5% CO₂ for 3 additional h. Cell viability was measured by recording the fluorescence signal of PrestoBlue (λ_exc/λ_em: 560/590 nm) using a Synergy H4 microplate reader (Biotek). Each experiment was performed in duplicates/triplicates from distinct samples.

Deiana M., Josse P., Dalinot C., Osmolovskyi A., Marqués P.S., Castán J.M., Abad Galán L., Allain M., Khrouz L., Maury O., Le Bahers T., Blanchard P., Dabos-Seignon S., Monnereau C., Sabouri N, & Cabanetos C. (2022). Site-selected thionated benzothioxanthene chromophores as heavy-atom-free small-molecule photosensitizers for photodynamic therapy. Communications Chemistry, 5, 142.

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Quantifying Cell Proliferation and Viability

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After cells are transfected with FBXL8-specific siRNA or control scrambled siRNA for 16 h, both Alamar blue and trypan blue dye exclusion were used to measure the cell proliferation. The cell growth was measured by Alamar blue. The metabolic activity of the cells was determined at 570 nm using a Synergy H4 microplate reader (BioTek) at time points indicated. For trypan blue dye exclusion, to assess total cell number, cells were scraped and resuspended in equal volumes of culture medium and trypan blue dye (0.4% solution; Gibco, Waltham, MA, USA) and counted using an improved Neubauer hemocytometer.

Chang S.C., Hsu W., Su E.C., Hung C.S, & Ding J.L. (2020). Human FBXL8 Is a Novel E3 Ligase Which Promotes BRCA Metastasis by Stimulating Pro-Tumorigenic Cytokines and Inhibiting Tumor Suppressors. Cancers, 12(8), 2210.

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Protein Extraction and DPP3 Activity Assay

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Total protein extracts were prepared from tail biopsies minced in 150 μL Tris–HCl, pH 7.8, and subjected to sonication and freeze-thawing cycles. For protein extracts from total bone, the tissue was frozen in liquid nitrogen and homogenized using aluminum beads and the TissueLyser II instrument (Qiagen, Hilden, Germany). Tissue debris were removed by centrifugation at 4°C. Protein concentration in the supernatants were estimated using the DC™ Protein Assay Kit II (Bio-Rad, Berkeley, CA, USA) following the manufacturer’s instructions, on a Synergy™ H4 Microplate Reader (BioTek Instruments, Inc., Winooski, VT, USA). DPP3 enzymatic activity was performed as reported,^{(31 (link))} with minor modifications. Briefly, 50 to 80 μg of protein extracts were assayed with 0.04 mM Arg-Arg-β-naphthylamide (Sigma-Aldrich, Saint Louis, MO, USA) in 250 μL Tris–HCl, pH 8.5, at room temperature (RT). The reaction was stopped by adding 50 μL of 2 M acetate buffer, pH 4.5, containing 10% Tween and 1.5 mg/mL Fast Blue B Salt (all chemicals from Sigma-Aldrich). The absorbance of the released product (β-naphthylamine) was measured at 530 nm, using SynergyTM H4 Microplate Reader. The enzymatic activity was expressed as percentage of the wild-type (WT) value.

Menale C., Robinson L.J., Palagano E., Rigoni R., Erreni M., Almarza A.J., Strina D., Mantero S., Lizier M., Forlino A., Besio R., Monari M., Vezzoni P., Cassani B., Blair H.C., Villa A, & Sobacchi C. (2019). Absence of Dipeptidyl Peptidase 3 Increases Oxidative Stress and Causes Bone Loss. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 34(11), 2133-2148.

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Assaying Complex I Activity and ATP Levels

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To assay complex I activity, wildtype and Cnhsf3∆ strains were cultured separately overnight and grown to an OD₆₀₀ of 0.8, then 100 μl of the resultant culture was mixed with 20 μl CellTiter 96® AQueous One Solution Reagent (Promega), which contains a novel tetrazolium compound, MTS. The mixture was placed in a 96-well assay plate and incubated at 30 or 40 °C for 1 h. NADH levels were determined by measuring absorbance at 490 nm using a MultiskanGO microplate reader (Thermo). All data were normalized using the number of colony-forming units (CFUs).
To monitor the ATP level, cells were prepared as described above. Strains were incubated at the indicated temperature for 30 min, then were disrupted using glass beads and a Bio-Spec bead beater for 3 rounds of 55 s each, resting on ice for 1 min between rounds. ATP levels were measured using the CellTiter-Glo Luminescent Cell Viability Assay (Beyotime) according to manufacturer instructions. The luminescent signal was measured using a Synergy H4 microplate reader (BioTek). All data were normalized to the protein concentration in the sample, which was determined using a MultiskanGO microplate reader (Thermo). Three independent experiments were performed, each including four technical replicates, and a representative data set is presented.

Gao X., Fu Y., Sun S., Gu T., Li Y., Sun T., Li H., Du W., Suo C., Li C., Gao Y., Meng Y., Ni Y., Yang S., Lan T., Sai S., Li J., Yu K., Wang P, & Ding C. (2022). Cryptococcal Hsf3 controls intramitochondrial ROS homeostasis by regulating the respiratory process. Nature Communications, 13, 5407.

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ROS-Glo Assay for HeLa Cells

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5 × 10³ HeLa cells/well were seeded in complete medium on 96 well-plates 24 h before the treatment with DBI. HeLa cells were washed (2 times) with live cell imaging solution (Molecular Probes™, Ref. No.: A14291DJ), then treated with DBI (1 μM) or DMSO (0.02% v/v) dissolved in live cell imaging solution and incubated for 90 min at 37°C in 5% CO₂. When required, the cells were irradiated with blue light using a LED light cube (Ex: 470/22 nm) operating at 30 mW cm⁻² for 4 min. After irradiation, ROS-Glo™ H₂O₂ kit (Promega) was used according to the manufacturer's instructions and the luminescence was recorded on a Synergy H4 microplate reader (Biotek).

Deiana M., Andrés Castán J.M., Josse P., Kahsay A., Sánchez D.P., Morice K., Gillet N., Ravindranath R., Patel A.K., Sengupta P., Obi I., Rodriguez-Marquez E., Khrouz L., Dumont E., Abad Galán L., Allain M., Walker B., Ahn H.S., Maury O., Blanchard P., Le Bahers T., Öhlund D., von Hofsten J., Monnereau C., Cabanetos C, & Sabouri N. (2023). A new G-quadruplex-specific photosensitizer inducing genome instability in cancer cells by triggering oxidative DNA damage and impeding replication fork progression. Nucleic Acids Research, 51(12), 6264-6285.

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Oxidase Activity of CNS in Buffers

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The oxidase activity of CNS at pH 4.0 and 7.0 was tested using organic dye tetramethylbenzidine (TMB) as substrate according to previous method [14 (link)]. In detail, 10 μg/ml CNS was mixed with 0.08 mM TMB in citrate buffer. Upon oxidation the TMB develop blue color which could be detected at 652 nm. The color change was recorded at interval of 30 s for 10 min using a Bio-TEK, Synergy H4 Microplate reader.

Dou C., Li J., He J., Luo F., Yu T., Dai Q., Chen Y., Xu J., Yang X, & Dong S. (2021). Bone-targeted pH-responsive cerium nanoparticles for anabolic therapy in osteoporosis. Bioactive Materials, 6(12), 4697-4706.

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Isolation of Kidney Mitochondria

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Kidneys were rinsed with Ringer’s solution, connective tissue was removed and immediately stored at 4 °C. Subsequently, the kidneys were homogenized in isolation medium A (220 mM mannitol, 70 mM sucrose, 5 mM TES, 0.1 mM EGTA, pH 7.3 with KOH) and centrifuged at 800× g (4 °C, 10 min). The supernatant was transferred to a clean centrifuge tube and centrifuged at 7200× g (4 °C, 10 min). The supernatant was stored as a cytosolic fraction and the pellet resuspended with medium A. The pellet fraction was centrifuged at 7200× g (4 °C, 10 min). Protein concentrations were determined by Bio-rad DC protein assay using a synergy H4 micro plate reader (Bio-Tek Instruments, Inc., Winooski, VT, USA) at absorbance 750 nm.

Star B.S., van der Slikke E.C., van Buiten A., Henning R.H, & Bouma H.R. (2023). The Novel Compound SUL-138 Counteracts Endothelial Cell and Kidney Dysfunction in Sepsis by Preserving Mitochondrial Function. International Journal of Molecular Sciences, 24(7), 6330.

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Cell Cytotoxicity Quantification with Cyquant

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The cytotoxic effects of LPS were quantified by Cyquant NF Cell Proliferation Assay Kit (ThermoFisher Scientific, #C35006) in cells, according to the manufacturer’s protocol. Cyquant binding to nuclear DNA was measured with the Synergy H4 micro plate reader (Bio-Tek) at excitation/emission of 497/520 nm.

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