El800 plate reader

Manufactured by Agilent Technologies

Sourced in United States

The EL800 plate reader is a compact and versatile instrument designed for absorbance-based microplate assays. It features a xenon flash lamp as the light source and can measure absorbance across a wide range of wavelengths. The EL800 is capable of processing microplates with 6 to 384 wells and supports various plate formats.

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Lab products found in correlation

Elisa, by Cayman Chemical (1 mentions) Elisa, by R&D Systems (1 mentions) Moloney murine leukemia virus reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Lightcycler 2.0 instrument, by Roche (1 mentions) Sybr green 1 master mix, by Roche (1 mentions) Elisa kit, by Hycult Biotech (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Quantikine elisa, by R&D Systems (1 mentions) Chromogenic lal endotoxin assay kit, by GenScript (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Cck 8, by Dojindo Laboratories (1 mentions) Assay buffer, by BioAssay Systems (1 mentions) Quantichrom gtpase assay kit, by BioAssay Systems (1 mentions) Celltiter 96 aqueous one mts assay, by Promega (1 mentions) 96 well tissue culture plate, by Corning (1 mentions) Oil red o staining, by Merck Group (1 mentions) Elx808, by Agilent Technologies (1 mentions) Elx405, by Agilent Technologies (1 mentions) Epmotion 5075lh, by Eppendorf (1 mentions) Hiv 1 p24 antigen elisa, by ZeptoMetrix (1 mentions)

29 protocols using el800 plate reader

Murine IL-33 and Cytokine Analysis

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Murine IL-33 protein content from BAL fluid, lung tissue lysates and peritoneal fluid were determined by ELISA per manufacturer’s instructions (R&D, Minneapolis, MN) and analyzed on a Biotek EL800 platereader (Winooski, VT). Detection of other murine cytokines (IL-4, IL-5, IL-13, IL-10, IL-6, IL-2, TNF, KC) from antigen restimulated leukocytes was performed using the Mouse V-PLEX Pro-inflammatory Panel 1 assay and the Mouse IL-13 Ultrasensitive assay per manufacturer’s instructions (Meso Scale Diagnostics, Rockville, MD) and plates were analyzed using a Sector S 600 plate reader. The number of samples assayed was based on the number of wells available on the plate and all data were reported. OVA-specific IgE and IgG1 from mouse plasma samples were assayed by ELISA (Cayman, Ann Arbor, MI).

Ogden H.L., Lai Y., Nolin J.D., An D., Frevert C.W., Gelb M.H., Altemeier W.A, & Hallstrand T.S. (2020). Secreted phospholipase A2 group X acts as an adjuvant for type-2 inflammation leading to an allergen-specific immune response in the lung. Journal of immunology (Baltimore, Md. : 1950), 204(12), 3097-3107.

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Mitochondrial Dehydrogenase Viability Assay

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Cell viability was also measured using MTT assay.^{17 (link)} Reduction of yellow MTT salt to a purple formazan crystals by mitochondrial dehydrogenases in viable cells was determined by measuring the light absorbance at 570 nm using EL× 800 plate reader (Bio-Tek Instruments Inc.)

Piotrowska A., Chmielewska M., Andrzejewski W., Dziegiel P, & Podhorska-Okolow M. (2020). Influence of Angiotensin II on cell viability and apoptosis in rat renal proximal tubular epithelial cells in in vitro studies. Journal of the Renin-Angiotensin-Aldosterone System: JRAAS, 21(3), 1470320320949850.

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Quantification of Inflammatory Markers

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Plasma levels of IL-33 and of sST2 were measured using the Quantikine ELISA (R&D Systems, Minneapolis, Minnesota, USA). To validate the ELISA measurements of IL-33, IL-33 mRNA expression was also measured by real time quantitative polymerase chain reaction (qPCR) in randomly selected patients in each sub-group (EHI, 9; CHI, 18; CHI-ART, 9; EC, 6 and UCs, 6). In brief, total RNA was extracted using RNeasy Mini kit (Qiagen, Hilden, Germany), and converted to cDNA using the Moloney murine leukemia virus Reverse Transcriptase (Life Technologies Inc., Burlington, Ontario, Canada). The cDNA was subjected to qPCR using LightCycler 2.0 Instrument – Roche, SYBR Green I master mix (Roche Diagnostics, Basel, Switzerland) as previously reported [20 (link)]. Intestinal-type fatty acid-binding protein (I-FABP), lipopolysaccharide (LPS), and soluble CD14 (sCD14) were measured in duplicate using commercially available ELISA kits from Hycult Biotech (Uden, the Netherlands), Cusabio (Wuhan, China), and R&D Systems, respectively. Optical densities were measured using the Biotek EL-800 plate reader (Winooski, Vermont, USA).

Mehraj V., Jenabian M.A., Ponte R., Lebouché B., Costiniuk C., Thomas R., Baril J.G., LeBlanc R., Cox J., Tremblay C, & Routy J.P. (2016). The plasma levels of soluble ST2 as a marker of gut mucosal damage in early HIV infection. AIDS (London, England), 30(10), 1617-1627.

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Quantification of Serum LPS Levels

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Serum LPS was quantified in triplicate from mice chronically treated with water (n = 4) or LPS (n = 4) according to the manufacturer’s instructions (Chromogenic LAL Endotoxin Assay Kit, #L00350, Genscript) using an EL800 plate reader (Bio-Tek Instruments) with absorbance at 540 nm. Positive control wells with serum containing three increasing concentrations of LPS were also run in triplicate. The limit of detection for the kit is 0.01–1 EU/ml.

Pittman D.W., Dong G., Brantly A.M., He L., Nelson T.S., Kogan S., Powell J, & McCluskey L.P. (2020). Behavioral and neurophysiological taste responses to sweet and salt are diminished in a model of subclinical intestinal inflammation. Scientific Reports, 10, 17611.

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MTT Assay for hBM MSC Cytotoxicity

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MTT (3-[4, 5-dimethylthiazol-2-yl]-2,5- diphenyltertra-zolium bromide) assay was used to investigate cytotoxic effects of various inducers on hBM MSC. The assay was performed at various time points of induction period (Day1, day3, day5, day7, day9 and day12) in all the four study groups with different inducers. Briefly, 1 × 10⁴ hBM MSC per well were plated onto flat-bottomed 48-well plates (Corning Glass Works, Corning, New York) with 500 μl of induction medium. At termination of different time points, the cells were subjected to 50 μl of MTT solution (5 mg/ml) for three hours at 37°C. Finally, 300 μl dimethylsulfoxide (Sigma, USA) was added to each well and incubated for 30 minutes at 37°C to dissolve all the formazan crystals. The coloured solution (200 µl) was transferred to 96 well plates and read at 570 nm using EL 800 plate reader (Biotek, USA) and recorded with Gen5 1.08.4 software (Biotek, USA). All the experiments were performed in triplicates for three samples each.

Nandy S.B., Mohanty S., Singh M., Behari M, & Airan B. (2014). Fibroblast Growth Factor-2 alone as an efficient inducer for differentiation of human bone marrow mesenchymal stem cells into dopaminergic neurons. Journal of Biomedical Science, 21(1), 83.

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Cytotoxicity of Retinol-Loaded Nanocarriers

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In vitro cytotoxicity of blank and retinol-loaded nanocarriers was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay on the HaCaT cell line (human keratinocytes, a kind gift from Pr. G. Weber, U1069 N2C, Tours) [41 (link)]. Cells were cultured in DMEM completed with 10% of SVF and 1% of a penicillin–streptomycin mixture. For the cytotoxicity study, cells were seeded in 96-well plates (10,000 cells/well) and incubated 24 h at 37 °C in a humidified atmosphere with 5% CO₂. Cells were then treated with increasing concentrations of blank or retinol-loaded nanocarriers (1.64 × 10⁻⁷ to 1.64 × 10⁻² mg/mL in retinol) during 24 h at 37 °C. MTT solution was added to each well (0.5 mg/mL) and the cells were incubated for further 4 h at 37 °C. Then, the supernatant was discarded and DMSO was added in each well to dissolve the formazan crystals. The absorbance A at 540 nm was measured with a microplate reader (Bio-tek EL800 plate reader). Results were expressed as viability percentages as function of the retinol concentration, and compared with Student’s t-test.

Van Gheluwe L., Buchy E., Chourpa I, & Munnier E. (2020). Three-Step Synthesis of a Redox-Responsive Blend of PEG–block–PLA and PLA and Application to the Nanoencapsulation of Retinol. Polymers, 12(10), 2350.

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Quantifying Flavonoid Content in Propolis

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The total flavonoid content (TFC) of the propolis extract was estimated using a previously described colorimetric method based on the formation of an aluminium chloride complex [13 ]. In an ELISA plate, 200 μL of the mixture was added in triplicate and the samples were incubated for 10 min in the dark at room temperature. A calibration curve was generated using different concentrations (1–100 μg/mL) of quercetin. The absorbance was read at 415 nm in a Bio-Tek EL800 plate reader (Bio-Tek) and the total flavonoid content was expressed as milligrams of quercetin equivalent per gram of extract (mg of QE/g of extract).

Rivera-Yañez N., Rodriguez-Canales M., Nieto-Yañez O., Jimenez-Estrada M., Ibarra-Barajas M., Canales-Martinez M.M, & Rodriguez-Monroy M.A. (2018). Hypoglycaemic and Antioxidant Effects of Propolis of Chihuahua in a Model of Experimental Diabetes. Evidence-based Complementary and Alternative Medicine : eCAM, 2018, 4360356.

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Cell Proliferation Assay with CCK-8

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Cell proliferation assays were performed using cell counting kit-8 (CCK-8; Dojindo Molecular Technologies, Kumamoto, Japan) according to the manufacturer’s instructions as described previously.⁵¹ (link) The indicated cells were seeded into 96-well plates at a density of 3 × 10³ cells per well, and the optical density (OD value) of each well at a wavelength of 450 nm at 72 h was detected using an EL×800 plate reader (BioTek Instruments, Winooski, VT, USA).

Lu Z., He Q., Liang J., Li W., Su Q., Chen Z., Wan Q., Zhou X., Cao L., Sun J., Wu Y., Liu L., Wu X., Hou J., Lian K, & Wang A. (2019). miR-31-5p Is a Potential Circulating Biomarker and Therapeutic Target for Oral Cancer. Molecular Therapy. Nucleic Acids, 16, 471-480.

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Malachite Green GTPase Activity Assay

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GTPase activity was determined with a malachite green assay [48 (link),53 (link)] using the QuantiChrom™ GTPase assay kit (BioAssay Systems). The assay was based on the formation of a stable green-coloured complex between the malachite green and P_i released during hydrolysis of GTP. Reactions were prepared to give a final volume of 150 μl, containing the assay buffer (BioAssay Systems), 10 mM MgSO₄, 1 mM GTP and 20–50 μg/ml of purified protein in 20 mM K-HEPES pH 7.0 buffer. The GTPase assay was performed at 37°C with continuous shaking after initiation of the reaction by addition of GTP (1 mM). Samples were taken at different time intervals over a total of 4 h. Reactions were terminated by addition of the colouring reagent (100 μl) and the A₆₃₀ values were read after 30 min using a BioTek EL800 plate reader. A standard curve was prepared simultaneously using phosphate dilutions (0–5 nM).

Seyedmohammad S., Fuentealba N.A., Marriott R.A., Goetze T.A., Edwardson J.M., Barrera N.P, & Venter H. (2016). Structural model of FeoB, the iron transporter from Pseudomonas aeruginosa, predicts a cysteine lined, GTP-gated pore. Bioscience Reports, 36(2), e00322.

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Quantifying Membrane Integrity via LDH Assay

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Membrane integrity of cells was characterized using LDH Assay (TOX-7, Sigma Aldrich, St. Louis, MO, USA) according to manufacturer’s protocol. NIH3T3 and MC3T3 cells at densities of 5,000 cells/well (15,625 cells/cm²) were seeded in a 96-well plate and incubated for 24 hours. After incubation, the media was replaced with the experimental media. After 24 hours treatment, the culture plates were centrifuged at a speed of 1200 rpm to remove cellular debris. 50 μl of the supernatant media was transferred to a new 96-well plate followed by addition of 100 μl LDH assay reagent to each of the wells. After incubation for 40 minutes in the dark, the absorbance was recorded at wavelength of 490 nm using a Biotek EL800 plate reader (Winooski, VT, USA). To the wells with 5000 viable cells, 10 μl lysis solution was added for 15 min (TOX-7, Sigma Aldrich, St. Louis, MO, USA) to lyse the cells, and these wells served as positive (dead) controls. Wells treated with experimental media of PPF polymer served as a baseline control. Wells containing blank media (without cells) served as a negative control. Sample size was n=6 for this assay. Fraction of dead cells was calculated using following equation:

Fraction of dead cells = A_{S} / A_{D}

where, A_S is the absorbance of each well after background subtraction, and A_D is the average absorbance of positive (dead) control after background subtraction.

Farshid B., Lalwani G, & Sitharaman B. (2014). In Vitro Cytocompatibility of One- and Two-Dimensional Nanostructure-Reinforced Biodegradable Polymeric Nanocomposites. Journal of biomedical materials research. Part A, 103(7), 2309-2321.

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