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Spectramax m2 reader

Manufactured by Molecular Devices
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The SpectraMax M2 reader is a multi-mode microplate reader designed for absorbance, fluorescence, and luminescence measurements. It features high-performance optics and a range of detection capabilities for a variety of assays and applications.

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Lab products found in correlation

Ebm 2, by Cambrex (2 mentions) Cck 8, by Dojindo Laboratories (2 mentions) Vegf a, by R&D Systems (1 mentions) 24 well plate, by Corning (1 mentions) Celltiter blue reagent, by Promega (1 mentions) Wst 1 solution, by Roche (1 mentions) H2dcfda, by Thermo Fisher Scientific (1 mentions) 96 well plate, by Corning (1 mentions) Platinum elisa kit, by Thermo Fisher Scientific (1 mentions) Rotina 380 r, by Hettich (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (1 mentions) Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Drabkin s solution, by Merck Group (1 mentions) Digital camera, by Nikon (1 mentions) Proteasome glo chymotrypsin like cell based assay, by Promega (1 mentions) Proteasome glo, by Promega (1 mentions) Proteasome glo assay, by Promega (1 mentions) Prestoblue, by Thermo Fisher Scientific (1 mentions)

20 protocols using spectramax m2 reader

1

Proteasome Activity Measurement in Cells

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Proteasome activity was measured using Proteasome-Glo (Promega), which detects chymotrypsin-like activity in cell-based assays. For proteasome activity assays, tissue/cell lysates were prepared in PIPES buffer (50 mM PIPES, 1 mM MgCl2, 50 mM NaCl, 2 mM EGTA, and 2 mM ATP) using glass dounce. 50 μg of protein extracts were aliquoted in duplicates to a black 96-well plate. Samples were assayed for proteasome activity with the Proteasome-Glo Chymotrypsin-like Cell-Based Assay (Promega) in a Spectramax M2 reader (Molecular Devices).
For PSMD5 KD experiments - HT-29 (5×103 cells per well) were seeded in a white 96 well plate. 24 hours later cells were transfected with control or PSMD5 siRNA (Dharmacon OnTarget pull of 5 different targeting RNAi’s) using Dharmafect1 transfection reagent as recommended by the manufacturer. Cells were analyzed 5–6 days post transfection using Proteasome-Glo assay (Promega). Luciferase values were normalized to cell viability as measured by PrestoBlue (Thermo Fisher Scientific) in a parallel plate. Samples were analyzed in a Spectramax M2 reader (Molecular Devices).
Levin A., Minis A., Lalazar G., Rodriguez J, & Steller H. (2018). PSMD5 inactivation promotes 26S proteasome assembly during colorectal tumor progression. Cancer research, 78(13), 3458-3468.
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2

Macrophage Differentiation and Inflammation

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THP-1 cells were harvested during the logarithmic growth phase and seeded into 96-well plates at 1 × 106 cells/well. Cells were treated with PMA (100 nM) for 24 h to induce their differentiation into resting M0 macrophages. Fresh complete culture medium was added and cultured allowed for 24 h after washing cells with phosphate-buffered saline. Then, differentiated THP-1 cells were stimulated with MSU (25, 50, 100, 150, 200, 300, 400 or 500 mg/ml) and treated with aurantiamide acetate (AA) at a final concentration of 0, 20, 60, 80 or 100 µM as well as colchicine (positive control, 5 µM) simultaneously for 24 h at 37 C (Xiao et al., 2019 (link)). After treatment, 10 μL of CCK-8 was added to each well, and the plates were incubated for an additional 1.5 h. Absorbance at 450 nm was measured (using 600 nm as a reference wavelength) on a SpectraMax™ M2 reader (Molecular Devices, Silicon Valley, CA, United States). The culture medium without cells was used as a blank. Cell survival was calculated as: absorbance/absorbance of control × 100%.
Ye X., Wu J., Zhang D., Lan Z., Yang S., Zhu J., Yang M., Gong Q, & Zhong L. (2021). How Aconiti Radix Cocta can Treat Gouty Arthritis Based on Systematic Pharmacology and UPLC-QTOF-MS/MS. Frontiers in Pharmacology, 12, 618844.
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3

THP-1 Macrophage Activation and Viability Assay

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THP-1 cells were harvested during the logarithmic growth phase and seeded into a 96-well plate at a density of 1x106 cells/well with a final volume of 100 μl. The cells were treated with PMA (100 nM) for 3 h to induce their differentiation into resting M0 macrophage. Fresh complete culture medium was added and cultured for 24 h after washing the cells with PBS solution. Differentiated THP-1 cells were then stimulated with MSU (100, 150, 200, 300, 400, and 500 μg/ml) with optimal concentration and treated with JSCBR extracts at final concentrations of 1, 2, 3, 4, and 5 mg/ml as well as colchicine (positive drug, 2 μg/ml) simultaneously at 37˚C for 24 h (Li et al., 2019 (link)). After treatment, 10 μl of CCK-8 was added to each well, and the plates were then incubated for additional 1.5 h. The absorbance at 450 nm was measured using 600 nm as a reference wavelength, which performed on a SpectraMax M2 reader (Molecular Devices, Silicon Valley, USA), and culture medium without cell was used as a blank. The experiments were performed in triplicate and repeated at least twice. The cell viability was calculated using the following formula:
where As, Ab, and Ac represent the absorbance difference of experimental, blank, and control groups, respectively.
Xiao N., Qu J., He S., Huang P., Qiao Y., Li G., Pan T., Sui H, & Zhang L. (2020). Exploring the Therapeutic Composition and Mechanism of Jiang-Suan-Chu-Bi Recipe on Gouty Arthritis Using an Integrated Approach Based on Chemical Profile, Network Pharmacology and Experimental Support Using Molecular Cell Biology. Frontiers in Pharmacology, 10, 1626.
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4

Cytotoxicity of Human Milk Oligosaccharides

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The cytotoxic effects of 3SG and 6SG were examined using an MTT labeling kit (Sigma-Aldrich). Briefly, HUVECs were cultured for 72 h in 24-well plates (Corning Inc.) in EBM-2 (Cambrex Inc.) containing 1% FBS at the indicated doses of HMOs and/or VEGF-A (50 ng/ml; R&D Systems Inc.). After removal of the culture medium, the cells were incubated with 300 μl of MTT (0.5 mg/ml) for 4 h at 37 °C in a 5% CO2 incubator. Formazan crystals in viable cells were dissolved in 300 μl of ethanol:DMSO (v/v, 1:1), and the absorbance in each well was measured at 540 nm using a SpectraMax M2 reader (Molecular Devices, Sunnyvale, CA).
Chung T.W., Kim E.Y., Choi H.J., Han C.W., Jang S.B., Kim K.J., Jin L., Koh Y.J, & Ha K.T. (2019). 6′-Sialylgalactose inhibits vascular endothelial growth factor receptor 2-mediated angiogenesis. Experimental & Molecular Medicine, 51(10), 120.
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5

Evaluating MSC Proliferation on PEG-Au Coatings

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PEG and PEG–Au solution was coated onto a culture dish to perform the following evaluation. First, 6 × 103 per well of MSCs were cultured in 96-well plates with pure PEG and PEG–Au 43.5 ppm coatings. The MSC alone group represented as the control. At 24, 48, and 72 h, an MTT (3-(4, 5-cimethylthiazol-2-yl)2, 5-diphenyltetrazolium bromide) solution (0.5 mg/mL) was added to each well and incubated for 3 h at 37 °C. Next, a Dimethyl-sulfoxide (DMSO) solution was added for 10 min to dissolve the crystals. The absorbance at 570 nm was detected by a SpectraMax M2 reader (Molecular Devices, San Jose, CA, USA).
Hung H.S., Yang Y.C., Kao W.C., Yeh C.A., Chang K.B., Tang C.M., Hsieh H.H, & Lee H.T. (2021). Evaluation of the Biocompatibility and Endothelial Differentiation Capacity of Mesenchymal Stem Cells by Polyethylene Glycol Nanogold Composites. Polymers, 13(23), 4265.
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6

CCK-8 Cell Viability Assay

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For CCK-8 assay, cells were counted and seeded at a density of 5,000 cells/well in 96-well plates containing complete culture medium. After treatment with 10 µL CCK-8 (Dojindo Laboratories, Kumamoto, Japan), absorbance was measured at 450 nm using a microplate reader (Spectra Max M2 reader; Molecular Devices LLC, Sunnyvale, CA, USA). Each experiment was performed in triplicate.
Yuan Y., Li S.L., Cao Y.L., Li J.J, & Wang Q.P. (2019). LKB1 suppresses glioma cell invasion via NF-κB/Snail signaling repression. OncoTargets and therapy, 12, 2451-2463.
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7

Evaluating BT Cytotoxicity in PR50-expressing Cell Lines

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On BT treatment, we replaced PR50-expressing SH-SY5Y and SK-N-DZ cells with fresh medium and treated them with serial dilutions of BT for 24 h, then collected cells for subsequent experiments. The toxicity of BT was confirmed using CellTiter-Blue® Reagent (Promega, Madison, WI, USA). The reagent was added directly to the culture medium for 2 h at 37 °C. Then, a SpectraMax M2 reader (Molecular Devices, Silicon Valley, CA, USA) was used to quantify the viability of the cells by fluorescent signal intensity (excitation: 560 nm, emission: 590 nm).
Fu R.H., Chen H.J, & Hong S.Y. (2023). Interaction of the C9orf72-Amyotrophic Lateral Sclerosis-Related Proline–Arginine Dipeptide Repeat Protein with the RNA-Binding Protein NOVA1 Causes Decreased Expression of UNC13A Due to Enhanced Inclusion of Cryptic Exons, Which Is Reversed by Betulin Treatment. Cells, 12(20), 2476.
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8

Cell Proliferation Assay with Growth Factors

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4500 cells with 100 μl of growth media were seeded in a
96-well plate in 10 replicates. After serum-starvation (0.5% FCS) for 26 hours,
cells were stimulated with 5–25 ng/ml of FGF2 or VEGFA,
10% FCS or 100μg/ml of insulin. After 72 hours,
10 μl of WST-1 solution (Roche Molecular Diagnostics)
was added for 2 hours. Absorption was measured
(A450nm-A690nm) by using a Spectramax M2 reader
(Molecular Devices).
Aimi F., Georgiopoulou S., Kalus I., Lehner F., Hegglin A., Limani P., Gomes de Lima V., A. Rüegg M., Hall M.N., Lindenblatt N., Haas E., Battegay E.J, & Humar R. (2015). Endothelial Rictor is crucial for midgestational development and sustained and extensive FGF2-induced neovascularization in the adult. Scientific Reports, 5, 17705.
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9

Quantifying Oxidative Stress in Fungal Spores

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The production of reactive oxygen species was measured as described previously (50 (link), 52 (link)), with minor changes. Briefly, 107 spores were incubated in 100 ml medium at 37°C for 24 h with shaking at 220 rpm. Then, 30 μM 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA; Invitrogen) was added to the medium and incubated at 37°C for 1.5 h. After this, the mycelia were harvested and washed three times with distilled water to remove extracellular H2DCFDA. The filtered mycelia were then ground in liquid nitrogen and suspended in PBS. The resulting supernatant was collected by centrifugation at 15,000 × g and 4°C for 10 min. Fluorescence was measured using a SpectraMax M2 reader (Molecular Devices, USA), with an excitation wavelength of 504 nm and an emission wavelength of 524 nm. The fluorescence intensity was normalized to the protein concentration of the sample, which was measured using a Thermo Coomassie protein assay kit.
Zhang Y., Wei W., Fan J., Jin C., Lu L, & Fang W. (2020). Aspergillus fumigatus Mitochondrial Acetyl Coenzyme A Acetyltransferase as an Antifungal Target. Applied and Environmental Microbiology, 86(7), e02986-19.
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10

MTT Assay for Cell Viability

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Cell viability was determined by MTT method. Five thousand cells/well were seeded in 96-well plates (Costar, Ann Arbor, MI). The next day, the HepG2 cells treated with rifampicin or DMSO were transfected with shRNA plasmid (0.2 μg) or pS-NC plasmid (0.2 μg). Drugs were added at increasing concentrations 48 h after transfection. Control cells were treated with an equal concentration of vehicle alone. 24 h later, the drug-containing medium was replaced by fresh medium and an aliquot of 20 μL of MTT (5 mg/mL) were added to each well. After incubation for 4 h at 37 °C, the supernatant was removed, and 150 μL of DMSO was added. The plates were vigorously shaken for 10 min to solubilize the MTT-formazan product. The absorbance was read using SpectraMax M2 reader (Molecular Devices, California, U.S.A) at a wavelength of 570 nm. Vehicle-treated cells were assigned a value of 100%13 , 15 . Each experiment was performed in triplicate wells for each drug concentration and carried out independently three times in different days. The cytotoxicity was evaluated with reference to the IC50 value. IC50 values were calculated from dose-response curves (i.e., cell survival versus drug concentration) obtained in multireplicated experiments using GraphPad Prism 5.0.
Xu S., Xiao Y., Li L., Yu L., Jiang H., Yu A, & Zeng S. (2014). Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression. Acta Pharmaceutica Sinica. B, 4(5), 350-357.
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