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Synergy 2 multi detection microplate reader

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The Synergy 2 Multi-Detection Microplate Reader is a versatile laboratory instrument designed for various detection modes. It provides consistent and reliable measurements for a wide range of applications in life science research, drug discovery, and analytical testing.

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Lab products found in correlation

Alamar blue, by Thermo Fisher Scientific (4 mentions) Revert aid reverse transcriptase, by Takara Bio (3 mentions) Trizol reagent, by Takara Bio (3 mentions) Dual luciferase assay system, by Promega (3 mentions) Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (3 mentions) Amplex red, by Thermo Fisher Scientific (2 mentions) Hek293 t cells, by Agilent Technologies (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Alamarblue, by Greiner (2 mentions) 96 well plate, by Greiner (2 mentions) Tritc dextran, by Merck Group (2 mentions) Tritc dextran, by Agilent Technologies (2 mentions) Transwell cell culture 8 μm inserts, by Merck Group (2 mentions) Bradford reagent, by Bio-Rad (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Pi3k elisa kit, by Echelon Biosciences (2 mentions) Pierce protein a g magnetic beads, by Thermo Fisher Scientific (2 mentions) Pgl3 basic luciferase vector, by Promega (1 mentions) Dual glo luciferase assay kit, by Promega (1 mentions)

108 protocols using synergy 2 multi detection microplate reader

1

Quantifying Bacterial Biofilm Formation

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Bacteria were scraped from the LB plate and resuspended in 500 μL PBS, diluted to 0.05 OD (595 nm) in 1 mL M9+CA (with 33.3Mm arabinose if needed). Five replicates of 100 μL of each sample were then transferred to a 96-well plate followed by 24 h incubation at 37°C. The following day, OD was measured using the Synergy 2 Multi-Detection Microplate Reader (BioTek) (595 nm). Planktonic bacteria were washed twice with 200 μL deuterium-depleted water (DDW), followed by the addition of 150 μL crystal violet (CV, Sigma-Aldrich Israel Ltd.) and left for incubation at room temperature (RT) for 15 min. The plate was then rinsed with water to remove crystal violet residues, and 200 μL absolute ethanol was added; the plate was incubated at RT for 15 min. One hundred μL elution from each well was transferred into a new 96-well plate, and the new plate was read in the Synergy 2 Multi-Detection Microplate Reader (BioTek) (OD 595 nm).
Shmidov E., Lebenthal-Loinger I., Roth S., Karako-Lampert S., Zander I., Shoshani S., Danielli A, & Banin E. (2022). PrrT/A, a Pseudomonas aeruginosa Bacterial Encoded Toxin-Antitoxin System Involved in Prophage Regulation and Biofilm Formation. Microbiology Spectrum, 10(3), e01182-22.
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2

Quantifying Bacterial Biofilm Formation

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Bacteria were scraped from the LB plate and resuspended in 500 μL PBS, diluted to 0.05 OD (595 nm) in 1 mL M9+CA (with 33.3Mm arabinose if needed). Five replicates of 100 μL of each sample were then transferred to a 96-well plate followed by 24 h incubation at 37°C. The following day, OD was measured using the Synergy 2 Multi-Detection Microplate Reader (BioTek) (595 nm). Planktonic bacteria were washed twice with 200 μL deuterium-depleted water (DDW), followed by the addition of 150 μL crystal violet (CV, Sigma-Aldrich Israel Ltd.) and left for incubation at room temperature (RT) for 15 min. The plate was then rinsed with water to remove crystal violet residues, and 200 μL absolute ethanol was added; the plate was incubated at RT for 15 min. One hundred μL elution from each well was transferred into a new 96-well plate, and the new plate was read in the Synergy 2 Multi-Detection Microplate Reader (BioTek) (OD 595 nm).
Shmidov E., Lebenthal-Loinger I., Roth S., Karako-Lampert S., Zander I., Shoshani S., Danielli A, & Banin E. (2022). PrrT/A, a Pseudomonas aeruginosa Bacterial Encoded Toxin-Antitoxin System Involved in Prophage Regulation and Biofilm Formation. Microbiology Spectrum, 10(3), e01182-22.
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3

Cytotoxicity Evaluation of OZ-PM on Calu-3 Cells

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In vitro, OZ–PM cytotoxicity, compared to the drug free micelles, was tested by measuring membrane damage on Calu-3 cells using the MTT assay [68 (link)]. Calu-3 cells lines were supplied from the American Type Culture Collection (ATCC, Manassas, VA, USA). Calu-3 cells were cultured in 96-well plates at a density of 6 × 104/well for 24 h under 5% CO2, 90% relative humidity, and at 37 °C. Different concentrations of OZ–PM or empty micelles were added to Calu-3 cells and incubated for 48 h. Consequently, 20 µL of 5 mg/mL of MTT solution was added to the cultured media and incubated for 4 h at 37 °C. Cell toxicity was analyzed by measuring absorbance at 570 nm with Synergy 2 Multi-Detection Microplate Reader, BioTek Instruments Inc. This assay was replicated three times for each experiment. The inhibitory concentration (50%) was determined, and the results were expressed as mean ± standard deviation. The negative control was prepared by adding culture medium alone (100% proliferation).
Abo El-Enin H.A., Ahmed M.F., Naguib I.A., El-Far S.W., Ghoneim M.M., Alsalahat I, & Abdel-Bar H.M. (2022). Utilization of Polymeric Micelles as a Lucrative Platform for Efficient Brain Deposition of Olanzapine as an Antischizophrenic Drug via Intranasal Delivery. Pharmaceuticals, 15(2), 249.
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4

Cell Viability Measurement by Crystal Violet

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N2a cells were grown and treated as in the cytotoxicity assay. Cell viability was determined by a modification of the crystal violet assay [23 (link)], and the optical density was measured at 540 nm with a 690-nm reference filter in a Synergy2 Multi-Detection Microplate Reader (BioTek Instruments, Inc.).
Elmann A., Telerman A., Ofir R., Kashman Y, & Lazarov O. (2018). β-amyloid cytotoxicity is prevented by natural achillolide A. Journal of Natural Medicines, 72(3), 626-631.
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5

Cloning and Luciferase Assay of CD47 Regulatory Regions

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The fragment of the CD47 regulatory regions was cloned by PCR using human genomic DNA as a template. The fragment was then cloned into the luciferase reporter plasmid pGL3-Basic Luciferase Vector (Promega). Cells were transiently transfected with desired pGL3 basic-based constructs. Luciferase activity was measured using the Dual-Glo® Luciferase Assay kit (Promega) with a Synergy 2 multi-detection microplate reader (BioTek, VT).
Liu F., Jiang C.C., Yan X.G., Tseng H.Y., Wang C.Y., Zhang Y.Y., Yari H., La T., Farrelly M., Guo S.T., Thorne R.F., Jin L., Wang Q, & Zhang X.D. (2017). BRAF/MEK inhibitors promote CD47 expression that is reversible by ERK inhibition in melanoma. Oncotarget, 8(41), 69477-69492.
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6

DPPH Radical Scavenging Activity Assay

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The DPPH radical scavenging activity assay was carried out in a Biotek Synergy 2 Multi-Detection Microplate Reader (Biotek, Winooski, VT, USA) according to the procedure described by Chandrasekar et al. [37 (link)] with minor modifications. All the experiments were run using a 400 uL 96-well plate. In brief, a series of samples with various concentrations in methanol were prepared, and then 50 uL of each sample solution was mixed with 50 uL of 0.2 mM DPPH solution freshly prepared in methanol. Methanol and l-ascorbic acid were used as the negative and positive control, respectively. After incubation for 30 min at room temperature in the dark, the absorbance of reactant was measured at 517 nm. The measurements of DPPH radical scavenging activity were carried out in triplicate. The percent of radical scavenging activity was determined from the difference in absorbance (A) of DPPH between the negative control and samples.
Radical scavenging (%) =[Anegative control−AsampleAnegative control]×100
Ruan X., Yang L., Cui W.X., Zhang M.X., Li Z.H., Liu B, & Wang Q. (2016). Optimization of Supercritical Fluid Extraction of Total Alkaloids, Peimisine, Peimine and Peiminine from the Bulb of Fritillaria thunbergii Miq, and Evaluation of Antioxidant Activities of the Extracts. Materials, 9(7), 524.
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7

Dual Luciferase Transcriptional Assays

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Dual luciferase transcriptional activity assays were performed as previously described (Guan et al., 2014 (link)). Fragments corresponding to 1,600 bp upstream of the TSS of TaGlu-1Bx, TaGlu-1By, TaGlu-1Dx, TaGlu-1Dy, TaSuSy1, and TaSSIIa were PCR-amplified from CS genomic DNA and cloned into the pGreenII 0800-LUC vector as reporter plasmids (Hellens et al., 2005 (link)). The ORFs of TaNAC019 and TaGAMyb were cloned into the pHB vector as effectors. Young leaves of 4-week-old N. benthamiana plants were co-infiltrated with Agrobacterium (strain GV3101) harboring different combinations of these plasmids. Firefly luciferase activity derived from the TaGlu-1Bx (TaGlu-1Bxpro:LUC), TaGlu-1Dy (TaGlu-1Dypro:LUC), TaGlu-1By (TaGlu-1Bypro:LUC), TaGlu-1Dx (TaGlu-1Dxpro:LUC), TaSuSy1 (TaSuSy1pro:LUC), or TaSSIIa (TaSSIIapro:LUC) promoters and Renilla luciferase under the control of the 35S promoter (35Spro:REN) were quantified using the Dual-Luciferase Reporter Assay system (Promega) with a Synergy 2 Multi-Detection Microplate Reader (BioTek Instruments). Normalized data are presented as the ratio of luminescent signal intensity for reporter versus internal control reporter (35Spro:REN) from three independent biological samples.
Gao Y., An K., Guo W., Chen Y., Zhang R., Zhang X., Chang S., Rossi V., Jin F., Cao X., Xin M., Peng H., Hu Z., Guo W., Du J., Ni Z., Sun Q, & Yao Y. (2021). The endosperm-specific transcription factor TaNAC019 regulates glutenin and starch accumulation and its elite allele improves wheat grain quality. The Plant Cell, 33(3), 603-622.
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8

Caspase-3/7 Activation Kinetics in Virus-Infected Cells

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Purified PBMs, seeded at 5 × 106 cells/mL in black polystyrene Nunc 96-well plates (Thermo Fisher), were cultured in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% heat-inactivated fetal calf serum (FCS), 100 IU/mL penicillin, 100 µg/mL streptomycin, and 100 ng/mL porcine colony stimulating factor -1 (CSF1) (Roslin Technologies) for 3 d. Prior to infection with the indicated virus at 0.25 MOI, the cells were loaded with 2 µM CellEvent Caspase-3/7 Green Detection Reagent (Thermo Fisher Scientific), according to the manufacturer’s instructions for kinetic assays and fluorescence in the live cells, indicative of caspase activity, recorded using a Synergy2 Multi-Detection Microplate Reader (BioTek) at the indicated time points post-infection.
Reis A.L., Rathakrishnan A., Goulding L.V., Barber C., Goatley L.C, & Dixon L.K. (2023). Deletion of the gene for the African swine fever virus BCL-2 family member A179L increases virus uptake and apoptosis but decreases virus spread in macrophages and reduces virulence in pigs. Journal of Virology, 97(10), e01106-23.
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9

ROS Measurement in N2a Cells

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N2a cells were grown and re-plated as in the cytotoxicity assay, using 1% instead of 5% FBS. After 24 h the cells were treated with 20 µM DCF-DA for 30 min at 37 °C. Following incubation, the cultures were rinsed with phosphate-buffered saline, and fresh medium was added to the cells. The ROS levels before and after treatment with achillolide A and Aβ were determined according to fluorescence (excitation at 485 nm and emission at 520 nm) in a Synergy2 Multi-Detection Microplate Reader (BioTek Instruments, Inc.).
Elmann A., Telerman A., Ofir R., Kashman Y, & Lazarov O. (2018). β-amyloid cytotoxicity is prevented by natural achillolide A. Journal of Natural Medicines, 72(3), 626-631.
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10

Quantifying NRF2 Activation in 3-NBA Metabolism

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The fluorescence of 3-ABA was used to measure the nitroreduction of nonfluorescent 3-NBA in cell culture as published previously.10 (link) To assess the contribution of NRF2 signaling in the metabolic activation of 3-NBA, experiments were conducted in various A549 cell lines (A549 wt, A549 NRF2-Het, and A549 NRF2-KO). Assays for 3-ABA formation in A549 cell lines were performed in 96-well plates 24 h (±2 h) after seeding of 1 × 104 A549 cells per well. After allowing 24 h for cells to attach, they were treated with 3-NBA in 0.1% DMSO in phenol red-free DMEM/F-12 (Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12). Formation of 3-ABA was measured (λex 520 nm, λem 650 nm) with a Synergy 2 multidetection microplate reader (Biotek Instruments Inc., Winooski, VT) and quantified by using calibration curves constructed with 3-ABA in phenol red-free DMEM/F-12 media.
Measurement of 3-ABA formation in HBEC3-KT cells was performed in 96-well plates by plating 2 × 104 cells which were allowed to recover for 24 h to allow them to attach, before they were treated with NRF2 activators (2.5–100 nM CDDO-Im or 0.625–10 μM SFN) or vehicle control (0.05% DMSO) for 48 h. Media was then replaced with fresh phenol red-free K-SFM containing 1.25–10 μM 3-NBA in 0.1% DMSO. Formation of 3-ABA (pmol) was quantified by using calibration curves constructed with 3-ABA in phenol red-free K-SFM.
Murray J.R., de la Vega L., Hayes J.D., Duan L, & Penning T.M. (2019). Induction of the Antioxidant Response by the Transcription Factor NRF2 Increases Bioactivation of the Mutagenic Air Pollutant 3-Nitrobenzanthrone in Human Lung Cells. Chemical research in toxicology, 32(12), 2538-2551.
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