Multiskan microplate spectrophotometer

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Multiskan microplate spectrophotometer is a laboratory instrument designed to measure the absorbance of samples in microplates. It provides accurate and reliable optical density measurements for a wide range of applications in life science research and clinical diagnostics.

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Lab products found in correlation

Eia ria plate, by Corning (2 mentions) Tmb solution, by Thermo Fisher Scientific (2 mentions) Rabbit anti human igg hrp, by Agilent Technologies (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (2 mentions) Powerup sybr green master mix, by Thermo Fisher Scientific (2 mentions) Hacat cells, by Thermo Fisher Scientific (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions) Maxisorp microtiter plate, by Thermo Fisher Scientific (1 mentions) Sars cov 2 spike s1 s2 ecd, by Sino Biological (1 mentions) Spike rbd, by Sino Biological (1 mentions) Diving pam, by Walz (1 mentions) Caspase3 activity assay kit, by Elabscience (1 mentions) Human tnf α standard abts elisa development kit, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin bsa, by GE Healthcare (1 mentions) Trichloroacetic acid tca, by Merck Group (1 mentions) Acetic acid, by Merck Group (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) E coli serotype 0111 b4, by Merck Group (1 mentions) Abi prism 7300 sequence detection system, by Thermo Fisher Scientific (1 mentions) Iscripttm cdna synthesis kit, by Bio-Rad (1 mentions)

31 protocols using multiskan microplate spectrophotometer

Cytokine Measurement in Plasma

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The levels of IL-6 and IL-10 were measured in plasma using the immunoenzymatic method called enzyme-linked immunosorbent assay (ELISA) (BD Bioscience, San Diego, CA, USA) in accordance with the manufacturer's instructions. Readings were performed using a Multiskan microplate spectrophotometer (Thermo LabSystems, Milford, MA, USA) at a 450 nm wavelength. The range of detection of the IL-6 kit used was 4.68 to 300 pg/mL, while the range of the IL-10 kit was from 7.8 to 500 pg/mL.
The TNF-α cytokine was measured using the Human TNF-α Standard ABTS ELISA Development Kit (Peprotech, Cranbury, NJ, USA) in accordance with the manufacturer's instructions. After 25 minutes, optical density was read using the Multiskan microplate spectrophotometer (Thermo Labsystems, USA) at 405 nm, using the 620 nm filter as reference wavelength. The detection range of the test was 62.5 to 4,000 pg/mL.

de Souza J.N., de Castro F.D., de Souza C.L., El Cheikh M.R., Ramos H.V., da Fonseca S.G, & Costa C.C. (2021). Is There a Difference between the Preoperative and Postoperative Serum Levels of Interleukin-6 and Tumor Necrosis Factor-α in Children Submitted to Adenotonsillectomy?. International Archives of Otorhinolaryngology, 26(2), e208-e212.

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Plasma HA-Specific IgG Quantification

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Plasma‐specific HA IgG was measured by ELISA. Briefly, EIA/RIA plates (Costar, St Louis, MO) were coated with HA antigen (2 μg mL^–1), blocked with 2% BSA and then incubated with diluted plasma samples (1:50). HA‐specific IgG was detected by adding rabbit anti‐human IgG HRP (Dako, Glostrup, Denmark). ELISA plates were developed using TMB solution (Life Technologies, Carlsbad, CA), and the reaction was stopped with 1 m HCl. Absorbance (OD450 nm) was measured using a Multiskan Microplate Spectrophotometer (Thermo Fisher). Serial dilutions of recombinant human IgG in separate wells on the same plate were performed for quantification of specific IgG.

Hartley G.E., Edwards E.S., Bosco J.J., Ojaimi S., Stirling R.G., Cameron P.U., Flanagan K., Plebanski M., Hogarth P.M., O’Hehir R.E, & van Zelm M.C. (2020). Influenza‐specific IgG1+ memory B‐cell numbers increase upon booster vaccination in healthy adults but not in patients with predominantly antibody deficiency. Clinical & Translational Immunology, 9(10), e1199.

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Cytotoxicity Evaluation of Orobol and Bentonite Composites

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The in vitro cytotoxicities of orobol, Bt-PC and Bt-PC@Orobol (2:2:1) composite formulation were evaluated in human keratinocyte (HaCaT) cells (Thermo Fisher Scientific, Massachusetts, USA). Briefly, cells were seeded on 96-well plates at 1.0 × 10⁴ cells/100 µL in complete Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 0.5% penicillin‒streptomycin. After applying various concentrations of orobol solution (3 × 10⁻⁴ ~ 3 × 10¹ µg/mL) or bentonite composite formulation (Bt-PC or Bt-PC@Orobol) at 1 × 10⁻³ ~ 3 × 10² µg/mL, the cells were incubated for 24 h or 48 h at 37 °C. Then, the cytotoxicity was assessed by a 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay (CellTiter 96 Aqueous One Solution Cell Proliferation Assay, Promega, Milan, Italy) according to the manufacturer’s instructions. The absorbance was read in a microplate reader (Multiskan Microplate Spectrophotometer; Thermo Fisher Scientific, Massachusetts, USA) at 492 nm.

Nguyen D.T., Kim M.H., Yu N.Y., Baek M.J., Kang K.S., Lee K.W, & Kim D.D. (2022). Combined Orobol-Bentonite Composite Formulation for Effective Topical Skin Targeted Therapy in Mouse Model. International Journal of Nanomedicine, 17, 6513-6525.

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PEG and COVID-19 Antibody Assays

Cited 1 time

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Anti-PEG IgG and anti-PEG IgM antibodies were detected with Human Anti-PEG IgG and IgM ELISA kit (PEGG-20 and PEGM-20, respectively; Gentaur, Kampenhout, Belgium). The kits were used according to the manufacturer’s instructions. All samples were run in duplicated and diluted 1:20. Optical density was measured for both assays at 450 nm and 620 nm (the latter for background subtraction) using a Multiskan Microplate Spectrophotometer (Thermo Scientific, Waltham, MA, USA). The antibody levels were calculated using the calibration curve built with the standards according to the protocol provided by the manufacturer. Results are reported as arbitrary units per unit volume (AU/mL).
Anti-spike and RBD IgG response was tested on Maxisorp microtiter plates (Nunc, Denmark) coated with recombinant SARS-CoV-2 Spike S1 + S2 ECD or Spike-RBD (all from Sino Biological, Chesterbrook, PA, USA), as previously reported [30 (link)].

Guerrini G., Gioria S., Sauer A.V., Lucchesi S., Montagnani F., Pastore G., Ciabattini A., Medaglini D, & Calzolai L. (2022). Monitoring Anti-PEG Antibodies Level upon Repeated Lipid Nanoparticle-Based COVID-19 Vaccine Administration. International Journal of Molecular Sciences, 23(16), 8838.

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Quantifying Zooxanthellae and Photosynthetic Yield

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Zooxanthellae cells were counted in triplicate under a light microscope using a hemocytometer in a 1:10 dilution. Chlorophyll was extracted using 90% acetone for 15 hr incubation at 4 °C and concentrations were determined using a Multiskan microplate spectrophotometer (Thermo Fisher Scientific, USA) and the equations previously described by Jeffery and Humphrey^{64 (link)}. Maximal photosynthetic yield (F_v/F_m) was measured using a Diving-PAM (Walz, Germany) for each polyp. Polyps were measured approximately 90 min after sunset to ensure a proper dark acclimation state prior to the measurement, both prior to their exposure to the light treatments and at the end of the experiment. The “measuring light intensity” and “gain” values were adjusted to reach optimal signals followed by an auto-zero calibration.

Ben-Zvi O., Eyal G, & Loya Y. (2019). Response of fluorescence morphs of the mesophotic coral Euphyllia paradivisa to ultra-violet radiation. Scientific Reports, 9, 5245.

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SARS-CoV-2 Antibody Detection by ELISA

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EIA/RIA plates (Costar, St Louis, MO) were coated with 2 μg/ml recombinant SARS-CoV-2 NCP or RBD or with hemagglutinin (HA) from influenza A/Michigan/08/2015 (AM15) overnight at 4°C (28 (link)). Plates were subsequently blocked with 3% BSA in PBS and incubated with diluted plasma samples, 1:30 for RBD and NCP and 1:50 for AM15. Antigen-specific IgG was detected by adding rabbit anti-human IgG HRP (Dako, Glostrup, Denmark). ELISA plates were developed using TMB solution (Life Technologies, Carlsbad, CA) and the reaction was stopped with 1 M HCl. Absorbance (OD450nm) was measured using a Multiskan Microplate Spectrophotometer (Thermo Fisher). Serial dilutions of recombinant human IgG (in-house made human Rituximab) in separate wells on the same plate were performed for quantification of specific IgG.

Hartley G.E., Edwards E.S., Aui P.M., Varese N., Stojanovic S., McMahon J., Peleg A.Y., Boo I., Drummer H.E., Hogarth P.M., O’Hehir R.E, & van Zelm M.C. (2020). Rapid generation of durable B cell memory to SARS-CoV-2 spike and nucleocapsid proteins in COVID-19 and convalescence. Science Immunology, 5(54), eabf8891.

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Caspase-3 Activity Assay in Breast Cancer Cells

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The Caspase-3 activity in
breast cancer cells, MDA-MB-231 and MCF-7, was examined using the
Caspase 3 Activity Assay Kit (Elabscience, E-CK-A311A) following the
manufacturer’s protocol. Briefly, MDA-MB-231 and MCF-7 cells
were seeded in a six-well plate at a density of 2 × 10⁵ cells and incubated overnight for 70–80% confluency. Post-incubation,
the cells were treated with respective IC₅₀ value of the
EA extract of C. gloeosporioides for
24 h. Post-treatment, the cells was trypsinized and washed once with
cold 1X PBS to remove excess culture media and trypsin. Further, the
cells were lysed with lysis buffer (containing DTT) for 30 min at
4 °C and centrifuged at 12,000 rpm for 15 min at 4 °C. The
supernatant containing Caspase-3 protein was collected, and the reaction
mixture was prepared by adding the protein sample, reaction buffer,
and substrate Ac-DEVD-pNA at a final volume of 100 μL and incubated
at 37 °C for 4 h. Post-incubation, the absorbance was recorded
at 405 nm using a Multiskan microplate spectrophotometer (Thermo Fisher
Scientific, USA), and the fold change in the activity of Caspase-3
was calculated. The representative result was from three independent
experiments.

Rai N., Gupta P., Verma A., Tiwari R.K., Madhukar P., Kamble S.C., Kumar A., Kumar R., Singh S.K, & Gautam V. (2023). Ethyl Acetate Extract of Colletotrichum gloeosporioides Promotes Cytotoxicity and Apoptosis in Human Breast Cancer Cells. ACS Omega, 8(4), 3768-3784.

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Bambara Groundnut Malt Antioxidant Potential

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The Bambara groundnut speciality malts and their syrup free radical scavenging ability were determined using the DPPH radical (25 mg/L) in 70% methanol. Each of the samples was mixed with 0.275 mL DPPH solutions. The samples and standards were incubated at 37 °C for 30 min in the dark, and absorbance reactions were read at 517 nm. The Thermo Scientific MultiSkan microplate spectrophotometer was used for reading absorbance. The standard was Trolox, and results were expressed as µmole Trolox/g.

Adetokunboh A.H., Obilana A.O, & Jideani V.A. (2022). Physicochemical Characteristics of Bambara Groundnut Speciality Malts and Extract. Molecules, 27(14), 4332.

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In Vitro Cytotoxicity Assay Protocol

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In vitro cytotoxicity assay
was performed as described previously by Chowdhury et al.^{33 (link)} Briefly, the cells were cultured in 96-well
plates. After 24 h, the cells were treated with drugs or NPs or compounds
for specific period of time. Thereafter, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide) (SRL) was added to each treated and control well and the
cells were incubated for 4 h. Formazan crystals were solubilized in
dimethyl sulfoxide (DMSO), and readings were obtained at 570 nm with
a differential filter of 630 nm using a multiskan microplate spectrophotometer
(Thermo Scientific). The percentage of viable cells was calculated
using the following formula: viability (%) = (mean absorbance value
of drug-treated cells)/(mean absorbance value of control) × 100.
A concentration of 0.2% DMSO was found to be nontoxic and was used
for dissolving CDDP and used as a control in cytotoxicity experiments.

Fageria L., Pareek V., Dilip R.V., Bhargava A., Pasha S.S., Laskar I.R., Saini H., Dash S., Chowdhury R, & Panwar J. (2017). Biosynthesized Protein-Capped Silver Nanoparticles Induce ROS-Dependent Proapoptotic Signals and Prosurvival Autophagy in Cancer Cells. ACS Omega, 2(4), 1489-1504.

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Measurement of cyclic nucleotides

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Cell density was adjusted to 5 × 10⁴ cells ml⁻¹. If needed, samples were preincubated with agonist or inhibitor for 10 min at room temperature. Subsequently, samples were kept in the dark or illuminated with light (520 nm, 20 µW mm⁻²) for 5 min at room temperature and lysed either immediately or after 3 min in the dark with 0.1 M hydrogen chloride. Concentrations of cGMP or cAMP were measured by DetectX Direct cyclic GMP or AMP Enzyme Immunoassay Kit, respectively, and analysed by the optical density at 450 nm with a Thermo Scientific Multiskan microplate spectrophotometer.

Zhang Y., Benz P., Stehle D., Yang S., Kurz H., Feil S., Nagel G., Feil R., Gao S, & Bender M. (2022). Optogenetic manipulation of cyclic guanosine monophosphate to probe phosphodiesterase activities in megakaryocytes. Open Biology, 12(8), 220058.

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