Synergy h1

Manufactured by Synergy Software

Sourced in United States

The Synergy H1 is a compact and versatile microplate reader designed for a wide range of applications. It features high-performance detection capabilities, including absorbance, fluorescence, and luminescence measurements. The Synergy H1 provides researchers with reliable and accurate data to support their experimental needs.

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Lab products found in correlation

Prism 8, by GraphPad (2 mentions) Black 96 well microplate, by Thermo Fisher Scientific (2 mentions) Cck8 reaction solution, by Solarbio (2 mentions) Rlt tissue lysis buffer, by Qiagen (2 mentions) Cck 8 solution, by Dojindo Laboratories (1 mentions) Buthionine sulfoximine bso, by Merck Group (1 mentions) Hydrochloric acid hcl, by Carl Roth (1 mentions) 5 sulfosalicylic acid ssa, by Merck Group (1 mentions) Tissuelyser, by Qiagen (1 mentions) Sterile 96 well plate, by Greiner (1 mentions) Cck 8 solution, by Beyotime (1 mentions) Caspase 3 9 activity kit, by Beyotime (1 mentions) Oseltamivir carboxylate, by MedChemExpress (1 mentions) Oseltamivir, by MedChemExpress (1 mentions) Prism v9, by GraphPad (1 mentions) Munana, by Merck Group (1 mentions) Sod activity assay kit, by Merck Group (1 mentions) Atp disodium salt, by Merck Group (1 mentions) Celltiter glo reagent from celltiter glo cell viability kit, by Promega (1 mentions) Dcfh da, by Thermo Fisher Scientific (1 mentions)

181 protocols using synergy h1

Bacterial Membrane Permeability Assays

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Outer membrane permeabilization was measured using 1-N-phenyl-napthylamine (NPN) [20 (link)]. E. coli (OD₆₀₀ = 0.1) in phosphate/citrate buffer with or without LH12 were mixed with 10 μM NPN. The increase of fluorescence intensity was monitored at excitation and emission wavelengths of 350 and 420 nm using a Synergy H1 multi-mode microplate reader. The experiment was performed in triplicate and repeated three times.
Inner membrane permeabilization was measured using o-Nitrophenyl-β-D-galactopyranoside (ONPG) [21 (link)]. E. coli was grown in Luria-Bertani broth with 2% lactose. Bacteria (10⁸ CFU/mL) in phosphate/citrate buffer with or without LH12 were mixed with 30 mM ONPG. The increase of OD₄₂₀ values was monitored using a Synergy H1 multi-mode microplate reader. The experiment was performed in triplicate and repeated three times.

Jiang W., Xie Z., Huang S., Huang Q., Chen L., Gao X, & Lin Z. (2022). Targeting cariogenic pathogens and promoting competitiveness of commensal bacteria with a novel pH-responsive antimicrobial peptide. Journal of Oral Microbiology, 15(1), 2159375.

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Fluorescent Probe for Sulfane Sulfur

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SSP4 (Sulfane Sulfur Probe 4) is a fluorescent probe to detect sulfane sulfurs specifically. SSP4 itself is non-fluorescent, but it emits strong green fluorescence after it reacts with sulfane sulfurs. Reactions of thiosulfate with SSP4 were conducted by mixing 10 μM SSP4 with 50 mM, 100 mM, or 300 mM thiosulfate in 200 μL HEPES buffer (100 mM, pH 3.0). The mixture was incubated at room temperature for 30 min, and then the fluorescence was detected by using the Synergy H1 microplate reader. The excitation wavelength was set to 482 nm and the emission wavelength was set to 515 nm.
For analysis of thiosulfate decomposition, 1 M thiosulfate was diluted into 200 μL HEPES buffer (100 mM, pH 4.0) to a final concentration of 300 mM. 100 mM Arg, Thr, or Gly was added individually. At the same time, 10 μM of SSP4 was added to incubate with thiosulfate for 30 min, and fluorescence was detected every 30 s by using the Synergy H1 microplate reader.

Wang Q., Li H., Xia Y., Xun L, & Liu H. (2021). Saccharomyces cerevisiae Rhodanese RDL2 Uses the Arg Residue of the Active-Site Loop for Thiosulfate Decomposition. Antioxidants, 10(10), 1525.

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Quantifying Chemokines CXCL2 and CXCL5 in BALF

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Chemokines including CXCL2 and CXCL5 in BALF were determined using ELISA kits
(R&D System, Minneapolis, MN) according to the instructions. For mouse
CXCL2, a 96-well microplate was coated with 100 µL per well of the capture
antibody. The plate was sealed and incubated overnight at room temperature. The
plate was blocked by adding 300 µL of reagent diluent and incubated for 1 hour.
Hundred microliters of samples or standards was added and incubated for 2 hours.
Hundred microliters of the detection antibody was added and incubated for 2
hours. Hundred microliters of streptavidin-horseradish peroxidase was added and
incubated for 20 minutes. Hundred microliters of substrate solution was added
and incubated for 20 minutes. Fifty microliters of stop solution was added, and
the optical density was determined using a microplate reader (Synergy H1) set to
450 nm.
For mouse CXCL5 immunoassay, 50 µL of assay diluent RD1W was added to each well.
Fifty microliters of standards, control, or samples were added and incubated at
room temperature for 2 hours. After being aspirated and washed, 100 µL of mouse
CXCL5 conjugate was added and incubated for 2 hours. Hundred microliters of
substrate solution was added and incubated for 30 minutes. Hundred microliters
of stop solution was added and the optical density was determined using a
microplate reader (Synergy H1) set to 450 nm.

Ye Y., Pei L., Wu C, & Liu S. (2019). Protective Effect of Traditional Chinese Medicine Formula RP on Lung Microenvironment in Pre-Metastasis Stage of Breast Cancer. Integrative Cancer Therapies, 18, 1534735419876341.

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Measuring Oxidative Stress and Glutathione in T84 Cells

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To examine ROS, T84 cells were pretreated with 5 μM 2′,7′-Dichlorofluorescin diacetate (H₂DCFDA; Sigma Aldrich Cat# D6883) for 1 hr at 37°C, 5% CO₂. Cells were then washed gently 2x with PBS and treated with 10 µg/mL ER stressor tunicamycin, or 2 mM H₂0₂ in the presence or absence of various concentrations of B. dentium LDM4 conditioned media, 2 mM γ-glutamylcysteine, or IL-10 in DMEM. Cells were incubated with treatment conditions for 3 hr, washed 3x with PBS, and then H₂DCFDA fluorescence was examined in cells in PBS on a Synergy H1 plate reader at excitation 485 nm/emission 520 nm. ThiolTracker Violet (ThermoFisher #T10095), an intracellular thiol probe used to detect glutathione levels. T84 cells were incubated with B. dentium LDM4 conditioned media, 2 mM γ-glutamylcysteine, or IL-10 in DMEM for 3 hr at 37°C, 5% CO_2. Cells were then washed and incubated with 20 μM ThiolTracker Violet in PBS for 30 min at 37°C, 5% CO_2. After incubation, cells were washed and fluorescence was examined on a Synergy H1 plate reader at excitation 404 nm/emission 526 nm. γ-glutamylcysteine uptake was examined using fluorescein (see supplemental methods).

Engevik M.A., Herrmann B., Ruan W., Engevik A.C., Engevik K.A., Ihekweazu F., Shi Z., Luck B., Chang-Graham A.L., Esparza M., Venable S., Horvath T.D., Haidacher S.J., Hoch K.M., Haag A.M., Schady D.A., Hyser J.M., Spinler J.K, & Versalovic J. (2021). Bifidobacterium dentium-derived y-glutamylcysteine suppresses ER-mediated goblet cell stress and reduces TNBS-driven colonic inflammation. Gut Microbes, 13(1), 1902717.

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ABTS Radical Scavenging Assay for Antioxidants

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The ABTS (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)) radical activity is based on an antioxidant’s capacity to scavenge the ABTS’s oxidized state. For this, ABTS radical scavenging activity is performed in a 96-well microplate, following the method described by [51 (link)] with some modifications. For the assay, ABTS radical was prepared by the mixture solution of equal parts of ABTS (7 mM) with potassium persulfate (2.44 mM), and then the radical was generated after 16 h. Before performing the assay in the microplate, ABTS radical absorbance was adjusted to 0.70 (±0.02) at 734 nm. In each well, for 20 µL of mucus samples was added 180 µL of ABTS radical. Instead of the sample, ultrapure water was used as control, and seven calibration solutions using Trolox (25–175 µM). The mixture was incubated for 5 min at 30 °C, and the absorbance at 734 nm was measured with a Multidetection plate reader (Synergy H1, Winooski, VT, USA). All measurements were performed in duplicate. The final results were expressed in µmol of Trolox equivalents.

Fernandez Cunha M., Coscueta E.R., Brassesco M.E., Marques R., Neto J., Almada F., Gonçalves D, & Pintado M. (2023). Exploring Bioactivities and Peptide Content of Body Mucus from the Lusitanian Toadfish Halobatrachus didactylus. Molecules, 28(18), 6458.

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Fluorometric Lipase Inhibition Assay

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The inhibition of lipase activity was determined by measuring the release of 4‐methylumbelliferone from 4‐MUO using a fluorimetric method (Cha, Song, Kim, & Pan, 2012). The fluorescence (A_a) (excitation at 360 nm and emission at 460 nm) was measured using the Synergy H1 instruments. Orlistat was used as a positive control, and all samples were assayed in triplicate. The percentage of lipase inhibition activity was calculated by the following equation:

inhibition (%) = [1 - \frac{(A_{a} - A_{b})}{(A_{c} - A_{d})}] \times 100,

where A_a, A_c, A_b, and A_d are the values of fluorescence of the sample, control, blank, and blank control, respectively.

Cheng K., Dong W., Long Y., Zhao J., Hu R., Zhang Y, & Zhu K. (2019). Evaluation of the impact of different drying methods on the phenolic compounds, antioxidant activity, and in vitro digestion of green coffee beans. Food Science & Nutrition, 7(3), 1084-1095.

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Quantification of Cry1Ac Protein in Plant Tissues

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The enzyme-linked immunosorbent assay (ELISA) kit AP003 CRBS (EnviroLogix, Portland) was used for Cry1Ac protein quantification assays. Fresh samples from the leaf, stem and endosperm at the late filling stage were placed in liquid nitrogen and ground into a powder. Approximately 20 mg of powder was suspended in 500 μL of the extraction/dilution buffer, and then diluted to appropriate concentrations (diluted 300 times for leaf tissue and 50 times for stem and endosperm tissue). The assay was carried out following the manufacturer’s instructions (EnviroLogix). Optical density values of the samples were measured at the 450 nm wavelength using a multi-mode microplate reader (Synergy H1, USA), and the values were used to calculate the content of Cry1Ac protein.

Liu X., Zhang C., Li X, & Tu J. (2016). Pyramiding and evaluation of both a foreign Bacillus thuringiensis and a Lysine-rich protein gene in the elite indica rice 9311. Breeding Science, 66(4), 591-598.

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Evaluating CAP-2M4VP Nanoparticle Cytotoxicity

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The effect of the CAP-2M4VP nanoparticles on host cell viability was assayed using Int 407 cell lines, cultured in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin (w/v), and 1% l-glutamine using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The Int 407 cell lines were procured from National Centre for Cell Science (NCCS), Pune, India. About 1 × 10⁴ Int 407 cells/well were seeded in a 96-well microtiter plate and incubated at 37 °C in a CO₂ incubator for 24 h. After the incubation period, the cells were treated with various concentrations (100 μg/mL, 50 μg/mL, 25 μg/mL, 12.5 μg/mL, 6.25 μg/mL) of CAP-2M4VP and incubated for another 24 h at 37 °C in a CO₂ incubator. After 24 h, 1 mg/mL of MTT was added, and the microtiter plate was incubated at 37 °C for 3 h. Dimethyl sulfoxide (DMSO) was added to dissolve the formazan crystals, and absorbance was read at 570 nm using a microplate reader spectrophotometer (Synergy H1)^{20 (link)}.

Sarveswari H.B., Gupta K.K., Durai R, & Solomon A.P. (2023). Development of a smart pH-responsive nano-polymer drug, 2-methoxy-4-vinylphenol conjugate against the intestinal pathogen, Vibrio cholerae. Scientific Reports, 13, 1250.

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Cytotoxicity Assay for Primary Cells

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Primary cells (1×10⁵/well) were seeded in 96-well flat-bottomed plates and cultivated in RPMI-1640 medium supplemented with 10% FBS, 50 μg/mL gentamicin, 50 μM 2-mercaptoethanol, and IL-2 (10 ng/mL). Cells were treated with different experimental conditions and after 48 h, CCK-8 solution (10 μL, Dojindo Molecular Technologies, Japan) was added into each well for 3-h incubation. The light absorption value (OD value) under each condition was recorded at 450 nm on a Synergy H1 microplate reader (USA).

Wang Y., Tu S., Huang Y., Qin K, & Chen Z. (2022). MicroRNA-181a regulates Treg functions via TGF-β1/Smad axis in the spleen of mice with acute gouty arthritis induced by MSU crystals. Brazilian Journal of Medical and Biological Research, 55, e12002.

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Adherent Cell Imaging Assay

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Non-adherent DCs were removed, and adherent cells were washed twice with 500 µl PBS. Adherent cells were stained with Hoechst 33,342 (NucBlue reagent, 2 drops/ml) in R10H20 medium for 30 min at 37°C. Cells were washed twice and 1 ml Live Cell Imaging solution (140 mM NaCl, 2.5 mM KCl, 1.8 mM CaCl₂, 1.0 mM MgCl₂, 20 mM HEPES, pH 7.4) was added to the wells. Fluorescence was measured with a Synergy H1 plate reader (excitation 490 nm, emission 520 nm, bottom reading without lid, 50 data points per well). Pictures of adherent cells were taken on a brightfield microscope at ×4 magnification and images were processed with Fiji.

Tomasek K., Leithner A., Glatzova I., Lukesch M.S., Guet C.C, & Sixt M. (2022). Type 1 piliated uropathogenic Escherichia coli hijack the host immune response by binding to CD14. eLife, 11, e78995.

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