RAW264.7 cells were seeded in 96-well plates at a density of 6.25×103 cells per well and allowed to incubate overnight. The next day, the cells were treated with different concentrations of engelitin (0, 5, 10, 20, 40 and 60μM) or DMSO. After 2 and 3 days, CCK-8 reagent was added and incubated in the incubator for 2 h. The absorbance of the solution was measured at 450 nm using a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies Gmbh & Co. KG, Baden-Württemberg, Germany).
Tristar2 lb 942 multimode microplate reader
Manufactured by Berthold Technologies
Sourced in Germany
The TriStar2 LB 942 Multimode Microplate Reader is a versatile laboratory instrument designed for the measurement and analysis of various assays in microplate format. It can perform absorbance, fluorescence, and luminescence detection across multiple wavelengths.
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9 protocols using tristar2 lb 942 multimode microplate reader
1
Engelitin Cytotoxicity Assay in RAW264.7 Cells
2
Evaluating Olaparib and Vemurafenib/Cobimetinib Synergy
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Cells (4 × 103) were seeded into 96-well plates. The following day, the media was replaced with 100 μl of olaparib-containing media, at final concentrations of 0, 10, 20 and 30 μM, together with 500 nM vemurafenib and 500 nM cobimetinib (Euromedex), in technical triplicates. Control cells were treated with olaparib-containing media (at 0, 10, 20 and 30 μM final concentration) together with DMSO. At the end of the treatment, cell viability was measured by a WST-1 ATP-based assay (Roche, France) after 48 h of treatment. Briefly, WST-1 reagent was added to each well (10 μL per 100 μL of medium) and incubated at 37 °C for 1.5 h. The plates were then read at 450 nm on a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies, Germany). Cell sensitivity is represented relative to 0 μM olaparib-treated cells. Statistical significance of the experimental data was determined using two-way ANOVA in GraphPad (Prism10).
3
Osteoclastogenesis Stimulation Assay
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Seeding of the BMMs was performed in 96-well plates based on a density of 6×103 cells per well at a temperature of 37°C and a concentration of 5 percent CO2, then, they were allowed to incubate overnight in order to adhere to the plate walls, followed by treatment with a complete α -MEM medium containing 25 ng/mL M-CSF and increasing dosages of Iso (0,0.625, 1.25, 2.5 or 5 μM) to stimulate the cells for 48 h. Subsequently, the cells were subjected to another incubation for 2 h with CCK-8 solution (10 µl/well). Finally, a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies Gmbh & Co. KG, Baden-Württemberg, Germany) was utilized to analyze the absorbance value (OD) at 450 nm.
4
Evaluating BMM Viability under PRI
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To estimate the viability of BMMs under PRI conditions, the cell counting kit-8 (CCK-8) was used. Briefly, BMMs uniformly planted with a number of 8×103 cells/well in 96-well plates were treated by diverse concentrations of PRI (0.3125, 0.625 and 1.25 μM) for 48 hours. At last, 10 μL CCK-8 was applied to culture cells for further 2 hours and the absorbance at 450 nm was detected using a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies Gmbh & Co.KG, Baden-Württemberg, Germany).
5
Quantifying Cytokine Secretion in Co-Culture
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In co-cultured system, the supernatant was collected at 4 h, 24 h, 48 h and centrifuged (1000 × g, 5 min) to remove the cells. The levels of secreted TGF-β1and IL6 in the supernatant were measured according to the manual of the enzyme-linked immunosorbent assay (ELISA) kit(Human TGF-β1 elisa kit: Neobiosience, Shenzhen, China, EHC107b.96. AuthentiKine Human IL-6 ELISA Kit, Proteintech, USA. Human IL-1 beta ELISA Kit, Proteintech).The absorbance value at a wavelength of 450 nm was measured with a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies, Bad Wildbad, Germany).
6
Circadian Rhythms in Arabidopsis Protoplasts
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Seeds were sown in 1/2 MS-solid medium and stratified in darkness at 4 °C for 3 days, and germinated seedlings were entrained for 3 weeks under ND conditions at 22–23 °C. To examine circadian oscillations in protoplasts, mesophyll protoplasts were isolated from the lower epidermal layer of 3-week-old Arabidopsis seedlings. Whole seedlings were soaked in 10 mL of an enzyme solution (400 mM mannitol, 20 mM KCl, 20 mM MES-KOH [pH 5.7], 10 mM CaCl2, 1% Cellulase R10, 0.5% Macerozyme R10, and 0.1% bovine serum albumin) for 16 h in the dark. The isolated protoplasts were filtered through sterile 100 mm stainless mesh and resuspended in W5 solution. The pCCA1:LUC and p35S:ELF3 plasmids were prepared by PEG purification. Then, the recombinant constructs were transiently introduced into Arabidopsis protoplasts via PEG-mediated transformation [23 (link)].
Luminescence rhythms were monitored using the Tristar2 LB 942 Multimode Microplate Reader (Berthold Technologies, Wildbad, Germany). The circadian period was estimated using the Fast Fourier Transform-Nonlinear Least Squares (FFT-NLLS) suite of programs available in the Biodare2 software.
Luminescence rhythms were monitored using the Tristar2 LB 942 Multimode Microplate Reader (Berthold Technologies, Wildbad, Germany). The circadian period was estimated using the Fast Fourier Transform-Nonlinear Least Squares (FFT-NLLS) suite of programs available in the Biodare2 software.
7
Cytokine Measurement in HUVEC Supernatant
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The levels of IL-1β, IL-6, and TNF-α in HUVEC culture supernatant were measured according to the manufacturer's protocol (MultiSciences Biotech Co., Ltd., China). The absorbance value at a wavelength of 450 nm was measured with a TriStar2 LB 942 Multimode Microplate Reader (Berthold Technologies, Germany).
8
Cellular ATP Quantification via Luminescence
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Cellular ATP levels were measured using ATP determination kit (Molecular Probes/Life Technologies, Canada) by adding 10 μl cell lysate in 100 μl ATP determination master mix containing 25 mM Tricine buffer, pH 7.8, 5 mM MgSO4, 0.5 mM D-luciferin, 1.25 μg/mL firefly luciferase, 100 μM EDTA and 1 mM DTT. The luminescence intensity was measured using TriStar 2 LB 942 Multimode Microplate Reader, Berthold Technologies, Germany. Protein content was determined by Bradford assay in each assay for normalization.
9
Cellular ATP Quantification via Luciferase Assay
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The cellular ATP level was measured using ATP determina-tion kit (Molecular Probes/Life Technologies, Canada) by using 1:10 diluted cell lysate in ATP determination master mix (25 mM Tricine buffer, pH 7.8, 5 mM MgSO4, 0.5 mM D-luciferin, 1.25 μg/ml firefly luciferase, 100 μM EDTA and 1 mM DTT). The luminescence intensity was measured using TriStar 2 LB 942 Multimode Microplate Reader, Berthold Technologies, Germany. The protein content was determined by Bradford assay and normalized with obtained intensity.
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