Spectramax id3 multi mode microplate reader

Manufactured by Molecular Devices

Sourced in United States, China, Austria

The SpectraMax iD3 Multi-Mode Microplate Reader is a compact, high-performance laboratory instrument designed for versatile microplate-based assays. The device can measure absorbance, fluorescence, and luminescence in microplates, allowing researchers to conduct a wide range of bioanalytical experiments.

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Lab products found in correlation

Celltiter blue cell viability assay, by Promega (4 mentions) Adp glo kinase assay, by Promega (4 mentions) Dual luciferase reporter assay system, by Promega (3 mentions) Softmaxpro7.1setup software, by Molecular Devices (2 mentions) Ix73 fluorescence microscope, by Olympus (2 mentions) Celltiter blue reagent, by Promega (2 mentions) Pierce ldh cytotoxicity assay kit, by Thermo Fisher Scientific (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions) Ganciclovir, by Merck Group (1 mentions) Cell titer glow, by Promega (1 mentions) Incucyte s3 live cell analysis system, by Sartorius (1 mentions) 5 5 dithiobis 2 nitrobenzoic acid dtnb, by Merck Group (1 mentions) Acetyl coa, by Merck Group (1 mentions) Oxaloacetic acid, by Merck Group (1 mentions) Jetprime, by Polyplus Transfection (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Cell culture lysis buffer, by Promega (1 mentions) Luciferase assay substrate, by Promega (1 mentions)

Spelling variants of this product

Microplate reader (2507 citations) SpectraMax iD3 (704 citations) SpectraMax microplate reader (255 citations) SpectraMax iD3 microplate reader (117 citations) Multi-mode microplate reader (40 citations) SpectraMax iD3 Reader (11 citations) SpectraMax iD3 multi (3 citations)

103 protocols using spectramax id3 multi mode microplate reader

Cytotoxicity Evaluation of Cell Viability and LDH Release

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Cell viability was based on the ability of viable cells to convert soluble MTT [3-(4,5n-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide] (Sigma Aldrich) into an insoluble purple formazan. Cells were seeded in 96-well plates. Briefly, cells grown and differentiated as described above were incubated for 4 h at 37°C with MTT (0.5 mg/mL in Neurobasal medium without phenol red). Then, the MTT solution was removed and water-insoluble formazan was immediately dissolved in DMSO (100 μL per well). The amount of formazan was measured at 570 nm with SpectraMax iD3 Multi-Mode Microplate Reader with a version of SoftMaxPro7.1Setup Software (Molecular Devices, LLC., San Jose, CA, USA).
Cytotoxicity was determined by measuring the release of LDH using the Pierce LDH Cytotoxicity Assay Kit (Thermo Fisher Scientific) according to the manufacturer's instructions. The absorbance was measured at 490 nm and 680 nm by SpectraMax iD3 Multi-Mode Microplate Reader with a version of SoftMaxPro7.1Setup Software (Molecular Devices, LLC.). To determine LDH activity, we subtracted the 680 nm absorbance value (background) from the 490 nm absorbance value before calculation of % cytotoxicity [(LDH at 490 nm)—(LDH at 680 nm)]. The percentage of the LDH release was normalized to the condition with the least amount of cell death and divided by a maximum lysis control.

Litwiniuk A., Domańska A., Chmielowska M., Martyńska L., Bik W, & Kalisz M. (2020). The Effects of Alpha-Linolenic Acid on the Secretory Activity of Astrocytes and β Amyloid-Associated Neurodegeneration in Differentiated SH-SY5Y Cells: Alpha-Linolenic Acid Protects the SH-SY5Y cells against β Amyloid Toxicity. Oxidative Medicine and Cellular Longevity, 2020, 8908901.

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Pharmacokinetics of ELPs and Rapamycin Formulations

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Male NOD mice aged 14–16 weeks were injected intravenously with 150 μL of 200 μM rhodamine-labeled ELPs (n = 5). Twenty μL of blood was collected from a tail nick at 5 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, 48 h, and 72 h after injection and immediately diluted into 80 μL of heparinized PBS. The blood was centrifuged at 16,100 rcf for 10 min at 4 °C, the supernatant was loaded onto a 384-well plate, and fluorescence was read using a SpectraMax iD3Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA) with excitation and emission wavelengths of 542 nm and 585 nm, respectively. After subtracting autofluorescence measured from the plasma of noninjected mice, the fluorescence intensity values were converted to concentration using a linear standard curve.
In a separate study, male NOD mice aged 14–15 weeks old were injected with 150 μL of 150 μM free Rapa (formulated in 90% PBS + 5% PEG400 + 5% Tween20), AFRapa, and IBPAF-Rapa through tail vein. Plasma was collected by cardiac puncture when the mice were under general anesthesia and subjected to Liquid chromatography–mass spectrometry (LC–MS) to measure Rapa concentration. Details of LC–MS are as described in the SI.

Ju Y., Guo H., Yarber F., Edman M.C., Peddi S., Janga S.R., MacKay J.A, & Hamm-Alvarez S.F. (2019). Molecular Targeting of Immunosuppressants Using a Bifunctional Elastin-Like Polypeptide. Bioconjugate chemistry, 30(9), 2358-2372.

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Resorufin-based Cell Viability Assay

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After treatment with H₂O₂ and/or CM or CM-derived factors, the number of viable cells was determined by a resorufin-based assay using the commercially available CellTiter-blue® cell viability assay (G8080, Promega, Sunnyvale, CA), according to the manufacturer’s protocol. MNs were cultured in a 96-well plate at a cell density of 2 × 10⁴ and treated with H₂O₂ and/or CMs for 24 h before performing the viability assay. A SpectraMax iD3 Multi‐Mode Microplate Reader (Molecular Devices, Sunnyvale, CA) was used to measure fluorescence intensity (560/590 nm).

Kim D., Kim S., Sung A., Patel N., Wong N., Conboy M.J, & Conboy I.M. (2022). Autologous treatment for ALS with implication for broad neuroprotection. Translational Neurodegeneration, 11, 16.

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Cell Viability and Proliferation Assays

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Cell viability was analyzed using the AlamarBlue cell viability reagent (Thermo Fisher) or CellTiter-Glow (Promega) according to the manual. Fluorescence emission was recorded with a SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices). To determine whether introduced mutations into the HBEGF locus affect the proliferative capacity, we evaluated cell growth of HEK293 wild-type cells and the produced HBEGF-mutant sublines. To monitor proliferation curves, 2000 cells were seeded per well of a 96-well plate and cell confluence was recorded every 24 h for 7 days, using the Incucyte S3 live-cell analysis system (Essen BioScience). For the experiments with the HSV-TK safety switch in hiPSCs, ganciclovir (Sigma, SML2346) was included in the cell culture media at 0.1, 1, and 10 µM concentration for 3 days, followed by a 3-day recovery.

Li S., Akrap N., Cerboni S., Porritt M.J., Wimberger S., Lundin A., Möller C., Firth M., Gordon E., Lazovic B., Sieńska A., Pane L.S., Coelho M.A., Ciotta G., Pellegrini G., Sini M., Xu X., Mitra S., Bohlooly-Y M., Taylor B.J., Sienski G, & Maresca M. (2021). Universal toxin-based selection for precise genome engineering in human cells. Nature Communications, 12, 497.

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Citrate Synthase Activity Assay

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Citrate synthase activity was determined using a previously described protocol (36 (link)). Acetyl-CoA (0.3 mM, Sigma-Aldrich) and oxaloacetic acid (0.5 mM, Sigma-Aldrich) were used as substrates for citrate synthase and 5,5-dithiobis (2-nitrobenzoic acid) (DTNB, 0.1mM, Sigma-Aldrich) were used to generate color. The reaction was activated with 10 μg snap-freeze cell lysate in reaction buffer (0.1 M Tris-HCl, pH 8.00). The absorbance was measured at 412 nm using a SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices, Shanghai, China).

Ye X., Wei X., Liao J., Chen P., Li X., Chen Y., Yang Y., Zhao Q., Sun H., Pan L., Chen G., He X., Lyu J, & Fang H. (2021). 4-Hydroxyphenylpyruvate Dioxygenase-Like Protein Promotes Pancreatic Cancer Cell Progression and Is Associated With Glutamine-Mediated Redox Balance. Frontiers in Oncology, 10, 617190.

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ELISA Protocol for Bacterial Quantification

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The ELISA test was performed
according to a previously established protocol with minor modifications.^{40 (link)} First, 100 μL of capture antibody (PA1–7213,
2 μg/mL in Carbonate-Bicarbonate Buffer, pH 9.6) was aliquoted
into a 96-well ELISA plate (44–2404–21, MaxiSorp, Fisher
Scientific, Spain) and incubated at 4 °C overnight. Second, 200
μL of BSA (3% in PBS) was added into the wells and incubated
at 37 °C for 1 h, to prevent nonspecific fouling of the wells.
Third, 100 μL of bacterial samples was added and kept at room
temperature for 1h. The wells were washed three times with PBST between
each step, and then incubated with 100 μL of HRP-labeled detection
antibodies (HRP-dAb, 0.33 μg/mL ab20425, in PBS) for 1 h at
room temperature. Finally, the ELISA wells were washed 5 times with
PBST. Afterward, 100 μL of the TMB substrate solution was added
into each well and left for 10 min at room temperature. Finally, 50
μL of 2 M H₂SO₄ was added into the wells
and then the absorbance of each well was detected at 450 nm using
a SpectraMax iD3Multi-Mode Microplate Reader (Molecular Devices, USA).

Zhao L., Rosati G., Piper A., de Carvalho Castro e Silva C., Hu L., Yang Q., Della Pelle F., Alvarez-Diduk R.R, & Merkoçi A. (2023). Laser Reduced Graphene Oxide Electrode for Pathogenic Escherichia coli Detection. ACS Applied Materials & Interfaces, 15(7), 9024-9033.

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Dual Luciferase Assay for XBP1s Transcriptional Activity

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The expression construct used for luciferase-based reporter assays is pcDNA3.1 XBP1s (RefSeq accession no. NM_001079539.1), while the reporter construct used is pGL3-PHGDH promoter region with XBP1 binding site CCACGT. For dual luciferase reporter assays, 2 × 10⁴ HEK-293T cells were plated overnight in a 96-well plate and transfected with the indicated plasmids using jetPRIME (Polyplus-transfection) according to the manufacturer’s protocol. Briefly, 18 ng of reporter and 2 ng of Renilla plasmid were cotransfected with various ratios (wt:wt) of expression plasmids (reporter:expression plasmid = 1:1, 1:3, or 1:5) and pcDNA3.1, which was added to reach a total of 200 ng of plasmid/well. After 48 h, cells were washed with PBS and lysed in Passive Lysis Buffer according to the manufacturer's protocol (dual luciferase reporter assay system, catalog #E1960; Promega). Firefly and Renilla luciferase activities were measured in white-bottom 96-well plates using an automated luminometer (SpectraMax iD3 Multi-Mode Microplate Reader; Molecular Devices). The reporter activity (Firefly) was normalized to its own Renilla luciferase activity.

Chaudhary V., Ah Kioon M.D., Hwang S.M., Mishra B., Lakin K., Kirou K.A., Zhang-Sun J., Wiseman R.L., Spiera R.F., Crow M.K., Gordon J.K., Cubillos-Ruiz J.R, & Barrat F.J. (2022). Chronic activation of pDCs in autoimmunity is linked to dysregulated ER stress and metabolic responses. The Journal of Experimental Medicine, 219(11), e20221085.

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Fluorescent Assay for Small Molecule Detection

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Probes were dissolved in DMSO to obtain a 1 mM stock solution. CTAB was dissolved in ethanol to yield a 5 mM stock solution. Analytes were dissolved in MilliQ water, 50 mM PBS (pH 7.4), or THF to make 10 mM stock solutions. Buffer was added to a 4.5 mL vial, followed by surfactant (25 μM), SSP4 (5 μM), and other species to a final volume of 4 mL. Samples were vortexed briefly and incubated in the dark at room temperature (rt) for 20 min before 3 mL was added to a 1 cm quartz cuvette and measured on a Cary Eclipse fluorescence spectrophotometer (emission 514 nm; excitation 482 nm). Samples measured on a Molecular Devices SpectraMax iD3 Multi-Mode Microplate Reader (emission 525 nm; excitation 485 nm; integration time 400s, low PMT sensitivity, attenuation 1, read height 1.00 mm) followed the same general procedure as above, though with a final volume of 200 μL added to a black polystyrene, flat-bottomed clear 96-well plate.

Shieh M., Ni X., Xu S., Lindahl S.P., Yang M., Matsunaga T., Flaumenhaft R., Akaike T, & Xian M. (2022). Shining a light on SSP4: A comprehensive analysis and biological applications for the detection of sulfane sulfurs. Redox Biology, 56, 102433.

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Evaluation of KRGE Fractions on Tau Aggregation

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The ThT assay was performed to investigate the effect of KRGE and its fractions on the dissociation of preformed tau aggregates. To induce tau aggregation, purified K18 (1 mg/mL) in phosphate-buffered saline (pH 7.4) was incubated with 0.1 mg/mL heparin and 100 μM DTT at 37°C for 24 hours (Figure S4). KRGE and KRGE fractions, including the SF, NSF, and NFP, were added to the aggregation mixture after 21 hours of incubation. At the end of the incubation period, ThT (15 μM in 50 mM glycine buffer, pH 8.9) was added to a 96-well plate and incubated for 3 hours. The fluorescence intensity of tau-bound ThT was measured at Ex/Em = 440 nm/484 nm with a SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices).

Shin S.J., Park Y.H., Jeon S.G., Kim S., Nam Y., Oh S.M., Lee Y.Y, & Moon M. (2020). Red Ginseng Inhibits Tau Aggregation and Promotes Tau Dissociation In Vitro. Oxidative Medicine and Cellular Longevity, 2020, 7829842.

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Luminescence-based Kinase Inhibition Assay

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The compound inhibitory activity was monitored based on the luminescence measured with the SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA, USA), using the previously described method with the aid of ADP-Glo kinase assay (Promega, Walldorf, Germany)^{35 (link)}. The measurements were carried in 96-well plate in volume of 25 μl in 20 mM Tris–HCl buffer (pH 7.5), containing 100 ng of hCK2α (1 μl), 10 μM CK2 substrate peptide RRRDDDSDDD (Biaffin GmbH & Co KG), 10 μM ATP, 20 mM MgCl₂ and 2.5 μl of ligand serially diluted in DMSO. The reaction was initiated by adding the enzyme and kept going for 20 min at 30 °C. For each ligand the standard dose–response curve was globally fitted to at least four independent experiments: common IC₅₀ value was optimized for all experiments, while top and bottom luminescence asymptotes were fitted individually for each experiment.

Paprocki D., Winiewska-Szajewska M., Speina E., Kucharczyk R, & Poznański J. (2021). 5,6-diiodo-1H-benzotriazole: new TBBt analogue that minutely affects mitochondrial activity. Scientific Reports, 11, 23701.

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