Eon microplate spectrophotometer

Manufactured by Agilent Technologies

Sourced in United States, Germany, Brazil

The Eon Microplate Spectrophotometer is a laboratory instrument designed for the measurement of absorbance in microplate formats. It provides accurate and precise spectrophotometric analysis of various samples, including biochemical, cellular, and microbiological assays.

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Lab products found in correlation

Vacutainer, by BD (3 mentions) Bs 21, by Jeio Tech (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (2 mentions) Tmb substrate, by Thermo Fisher Scientific (2 mentions) Transcriptor first strand cdna synthesis kit, by Roche (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Bovine elisa kit, by Mercodia (2 mentions) Luria bertani medium, by Merck Group (1 mentions) Human dkk1 quantikine elisa kit, by R&D Systems (1 mentions) Af1096, by R&D Systems (1 mentions) Anti dkk1, by R&D Systems (1 mentions) Apotome microscope, by Zeiss (1 mentions) Hapt 11, by Life Diagnostics (1 mentions) Cell count reagent sf, by Nacalai Tesque (1 mentions) Clear flat bottom 96 well plate, by Corning (1 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) 96 well microplate, by Corning (1 mentions) Fastin elastin kit, by Biocolor (1 mentions)

Spelling variants of this product

Microplate spectrophotometer (506 citations) Eon spectrophotometer (33 citations) Eon™ High Performance Microplate Spectrophotometer (15 citations) EonTM Microplate Spectrophotometer (7 citations) Eon Microplate (4 citations)

164 protocols using eon microplate spectrophotometer

Measurement of α,β-Unsaturated Aldehydes in Frying Oil

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The α,β-unsaturated aldehyde content was measured according to the p-anisidine test, the American Oil Chemists’ Society official method (Tompkins and Perkins, 1999 (link)). The frying oil (1 g) was dissolved in 25 mL isooctane and used as a sample solution. The sample solution (5 mL) was mixed with 1 mL glacial acetic acid, and its absorbance (A₀) was measured at 350 nm (EON microplate spectrophotometer, BioTek Instruments). The sample solution (5 mL) and isooctane (5 mL) were separately taken in each test tube, and 1 mL of the p-anisidine solution (0.25 g p-anisidine/100 mL glacial acetic acid) was added to both test tubes. After incubation at 23°C in a water bath (BS-21, Jeio Tech Co., Ltd.) covered with a lid for 10 min, the absorbance (each A₁ and A₂) at 350 nm (EON microplate spectrophotometer, BioTek Instruments) was measured. Isooctane was used as a blank. p-AV was calculated with the following equation:

p -AV= \frac{25[1 {.2(A}_{1} {-A}_{2} {)-A}_{0}]}{m}

where m is the mass (g) of the frying oil sample. The TOTOX value, an indicator of TOTOX, was calculated as follows:

TOTOX=2×PV+ p -AV

Lee J, & Surh J. (2022). Effects of Sweet Potato Powder Selected Based on the Polar Paradox Hypothesis on Oil Oxidation in the Preparation of Deep-Fried Croquettes. Preventive Nutrition and Food Science, 27(2), 248-256.

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Antibacterial Efficacy of HPMA-PEP Constructs

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The antibacterial properties of HPMA-PEP constructs were tested against Acinetobacter baumanii (CCM 2355), Escherichia coli (CCM 4517), Staphylococcus epidermidis (CCM 2124), and Staphylococcus aureus (CCM 4516) (Czech Collection of Microorganisms, Brno, Czech Republic) in vitro. A single colony of the test microorganism was transferred into 5 mL of sterile broth (Luria-Bertani medium, Sigma) and cultivated overnight at 37 °C to prepare a bacterial test suspension in a double-concentrated Mueller-Hinton broth at a density equivalent to a 0.5 McFarland standard. Desired weight of lyophilized PEP or polymer-PEP was resuspended in sterile PBS and a concentration series was prepared. The bacterial suspension was then mixed with different concentrations of the PEP or polymer-PEP construct diluted in 1:1 in PBS and cultured at 35 °C. The bacterial growth was measured spectrophotometrically at 625 nm every 60 min for 20 h using an EON microplate spectrophotometer (Agilent Technologies, Santa Clara, CA, USA).
The MIC was determined as the lowest concentration of PEP or polymer-PEP construct without observable bacterial growth. MIC was calculated as the molar concentration of PEP. The MBCs were also evaluated by culturing the samples (25 µL in 5 replicates) at MIC on plate count agar at 37 °C. If the bacteria were absent after 24 h of incubation, it was termed MBC.

Pola R., Vícha M., Trousil J., Grosmanová E., Pechar M., Rumlerová A., Studenovský M., Kučerová E., Ulbrich P., Vokatá B, & Etrych T. (2023). Polymer-Antimicrobial Peptide Constructs with Tailored Drug-Release Behavior. Pharmaceutics, 15(2), 406.

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Quantitative DKK1 Measurement Protocols

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Secreted DKK1 levels were measured either by Western blot after immunoprecipitating DKK1 from the culture medium using anti-DKK1 (#AF1096; R&D Systems) or by commercially available ELISA assay (Human Dkk-1 Quantikine ELISA Kit, #DKK100B; R&D Systems) according to the manufacturer’s protocol. For DKK1 ELISA, absorbance was measured at 450 nm with a correction at 540 nm using the Eon microplate spectrophotometer (Agilent).

Kim H., Jang S, & Lee Y.S. (2022). The m6A(m)-independent role of FTO in regulating WNT signaling pathways. Life Science Alliance, 5(5), e202101250.

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Live Imaging of Intracellular Calcium

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In all, 1 × 10⁵ cells were grown on a glass coverslip in 35 mm dishes. For performing live imaging cells edges of a glass slide were covered with parafilm and coverslip containing cells was inverted and put on a glass slide. The space between coverslip and glass slide was filled with cell culture media to keep cells alive. Images of cells were taken with the help of ×63 objective on an ApoTome microscope (ZEISS). For each field of view, donor alone, acceptor alone, and FRET channel micrographs were captured. These confocal micrographs were analysed by FRET and Colocalization analyzer plugin in ImageJ. FRET/CFP normalised emission ratio was plotted against time using ggplot2 R package. For endpoint analysis of intracellular calcium levels, cells were cultured and infected in 96 well black assay plates. Fluorescence intensities were analysed using Eon Microplate Spectrophotometer (Agilent Technologies).

Kumar A., Mishra S., Kumar A., Raut A.A., Sato S., Takaoka A, & Kumar H. (2022). Essential role of Rnd1 in innate immunity during viral and bacterial infections. Cell Death & Disease, 13(6), 520.

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Serum Haptoglobin Concentration Measurement

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Serum haptoglobin (HPT) concentrations on 35 and 42 d were measured using a commercial kit (Life Diagnostics Inc., West Chester, PA, USA, catalog# HAPT-11) following the manufacturer’s protocol. Briefly, serum samples were diluted (1:50) first with the diluent of the kit. The diluted samples and the standards (100 µL) were transferred in duplicate to a 96-well plate and then incubated on an orbital shaker (150 rpm) at room temperature for 45 min. Then, 100 µL of a horseradish peroxidase conjugate was added to each well and incubated on the orbital shaker (150 rpm) at room temperature for 45 min. Tetramethylbenzidine substrate (100 µL) was dispensed to each well, and the plate was incubated on the orbital shaker (150 rpm) at room temperature for 20 min. After adding the stop solution (100 µL) to each well, the optical density was read at 450 nm by using an Eon microplate spectrophotometer (BioTek Instruments, Inc., Winooski, VT, USA). The samples were reanalyzed if the coefficient of variation (CV) across duplicates was >20%. The intra-assay CV was <10% for 90% of the samples, and the inter-assay CV was 2.0%. The detection limit of this assay was 4.0 ng/mL.

Wickramasinghe J., Kaya C.A., Beitz D, & Appuhamy R. (2022). Early Stepdown Weaning of Dairy Calves with Glutamine and Branched-Chain Amino Acid Supplementations. Animals : an Open Access Journal from MDPI, 12(12), 1474.

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Quantification of Alizarin Red and Oil Red

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Alizarin Red quantification was performed following a previously described method [24 (link)]. Oil Red quantification was also performed following previously described protocols [25 (link)]. Absorbance lecture of the plates was performed using an Eon Microplate Spectrophotometer (BioTek, Winooski, VT, USA).

García-Sánchez D., González-González A., García-García P., Reyes R., Pérez-Núñez M.I., Riancho J.A., Évora C., Rodríguez-Rey J.C, & Pérez-Campo F.M. (2021). Effective Osteogenic Priming of Mesenchymal Stem Cells through LNA-ASOs-Mediated Sfrp1 Gene Silencing. Pharmaceutics, 13(8), 1277.

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Bacterial Growth in Oxalate Assays

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Bacillus subtilis strain 168 (ATCC 23857) and Escherichia coli (UTI89) were routinely cultured at 37°C in Luria-Bertani (LB) broth (61 (link)). Growth in oxalate was assayed in 96-well plates prepared as dilutions of sodium oxalate (NaOx) in LB broth with stationary-phase bacteria added at a final dilution of 1/100. Plates were incubated for 24 h at 37°C with optical density (OD) readings every 30 min using an Eon microplate spectrophotometer (BioTek, Winooski, VT, USA). NaOx concentrations were selected based on physiologic intestinal oxalate concentrations reported in the literature (62 (link)). Growth curves were analyzed with the R package Growthcurver and GraphPad Prism (version 8.1.2) (63 (link)).
D. melanogaster flies were cultured following homogenization until day 10 of the survival assay to assess BS168 loads. D. melanogaster flies were surface sterilized with 70% ethanol and homogenized in 0.01 M phosphate-buffered saline (PBS) by using a motorized pestle. Homogenates were diluted up to 100-fold, plated onto LB agar, and then incubated aerobically at 37°C for 24 h. The characteristic colony morphology of BS168 was easily differentiated on LB, which is not a growth medium amenable to culturing the typical D. melanogaster microbiota (64 (link)).

Al K.F., Daisley B.A., Chanyi R.M., Bjazevic J., Razvi H., Reid G, & Burton J.P. (2020). Oxalate-Degrading Bacillus subtilis Mitigates Urolithiasis in a Drosophila melanogaster Model. mSphere, 5(5), e00498-20.

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Quantification of T Cell Cytokine Production

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IL-2 and IFN-γ production by T cells was detected using the BD Biosciences recommended ELISA protocol. CD4⁺ T cell production of IL-2 was captured from supernatants using rat anti-mouse IL-2 (JES6–1A12) and detected using biotinylated rat anti-mouse IL-2 (JES6–5H4) antibodies. Whereas CD8⁺ T cell production of IFN-γ was measured in supernatants using rat anti-mouse IFN-γ (R4–6A2) and biotinylated rat anti-mouse IFN-γ (XMG1.2) antibodies. Graded amounts of recombinant murine IL-2 and IFN-γ were included in the assays for generation of standard curves from which reported concentrations were extrapolated using the linear portion of the curve. The IL-2 and IFN-γ concentrations were acquired using a BioTek Eon microplate spectrophotometer (BioTek Instruments Inc.) and analyzed using Gen 5 software version 2.01.14.

Bradley J.H., Barwick S., Horn G.Q., Ullrich E., Best B., Arnold J.P, & Gregg R.K. (2019). Simulated microgravity-mediated reversion of murine lymphoma immune evasion. Scientific Reports, 9, 14623.

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Comprehensive Physiological Biomarker Analysis

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Blood lactate concentrations (mM) were analyzed in real-time from plasma using an automated analyzer (Analox GM7 enzymatic metabolite analyzer; Analox Instruments USA, Lunenburg, MA, USA). Plasma concentrations of TAC, total testosterone, ferric-reducing ability of plasma (FRAP), and thiobarbituric reactive substances (TBARS), as well as serum concentrations of creatine kinase (CK), cortisol and myoglobin, were assayed using commercially available enzyme-linked immunosorbent assay kits. The TAC was assessed as the combined antioxidant ability of the sample to prevent the oxidation of 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) and is expressed in Trolox equivalents. Assay absorbance was read according to manufacturer specifications on a BioTek Eon Microplate Spectrophotometer (BioTek Instruments, Inc., Winooska, VT, USA). To eliminate interassay variance, all samples for a particular assay remained frozen until analysis, were thawed only once, and were measured in duplicate by a single technician. The coefficient of variation for each assay was 4.7% for TAC, 1.5% for lactate, 7.6% for myoglobin, 3.7% for CK, 4.3% for testosterone, 3.4% for cortisol, 14.4% for FRAP, and 6.5% for TBARS. All assay procedures followed the manufacturers' guidelines.

Beyer K.S., Stout J.R., Fukuda D.H., Jajtner A.R., Townsend J.R., Church D.D., Wang R., Riffe J.J., Muddle T.W., Herrlinger K.A, & Hoffman J.R. (2017). Impact of Polyphenol Supplementation on Acute and Chronic Response to Resistance Training. Journal of Strength and Conditioning Research, 31(11), 2945-2954.

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Determination of Frying Oil Peroxide Value

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The PV of the frying oils was determined according to the ferric thiocyanate method, which is based on the ability of peroxides to oxidize ferrous ions (Fe²⁺) to ferric ions (Fe³⁺; Chapman and Mackay, 1949 (link)). The FeCl₂ solution was prepared by adding 0.0328 M BaCl₂ solution slowly with stirring to 0.036 M FeSO₄-7H₂O solution and precipitating excess BaSO₄ with 10 N HCl (1 mL). The frying oil (0.01 g) was dissolved in 3 mL methanol/butanol (2:1, v/v). The sample solution was sequentially added with 15 μL ammonium thiocyanate (7.5 g/25 mL in water) and 15 μL FeCl₂ solution and vortexed thoroughly. After allowing the mixture to react at room temperature in the dark for 20 min, its absorbance was measured at 510 nm (EON microplate spectrophotometer, BioTek Instruments). The PV of the oil sample was calculated using a standard curve (R²=0.9934) constructed with cumene hydroperoxide.

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