NK92 or primary NK cells were incubated for 12 h in NK92 culture media supplemented with 0.1% BSA instead of 12.5% FBS and 12.5% horse serum. After washing, cells were incubated with various concentrations of peptides (control, cov1, cov2) for 2 h, and then NK cytotoxicity was evaluated by a calcein-AM release assay [30] (link). Briefly, target cells were labeled with calcein-AM (Invitrogen, Carlsbad, CA, USA) for 1 h. Then, calcein-labeled target cells (1 × 104 cells per well) and serially diluted NK cells were co-cultured in 96-well round-bottom plates for 4 h. “Maximum release” was simulated by adding 2% Triton X-100 to the target cells, and “spontaneous release” was simulated by adding culture medium to the target cells. The calcein released into the supernatant was measured using a multi-mode microplate reader (Molecular Devices, San Jose, CA, USA). The percent specific lysis was calculated according to the formula ((test release-spontaneous release)/(maximum release-spontaneous release)) × 100.
Multi mode microplate reader
Manufactured by Molecular Devices
Sourced in United States, China
The Multi-mode microplate reader is a versatile lab equipment designed to measure various types of assays in a microplate format. It is capable of performing absorbance, fluorescence, and luminescence measurements across multiple wavelengths.
Automatically generated - may contain errors
Lab products found in correlation
Calcein am, by Thermo Fisher Scientific (3 mentions)
Calcein, by Thermo Fisher Scientific (3 mentions)
Calcein, by Molecular Devices (3 mentions)
E2129, by Merck Group (1 mentions)
Mtt assay, by Merck Group (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Epiquik nuclear extraction kit, by Epigentek (1 mentions)
Fluor de lys hdac fluorometric activity assay kit, by Enzo Life Sciences (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Ldh cytotoxicity detection kit, by Takara Bio (1 mentions)
Fluorescence microscopy, by Zeiss (1 mentions)
Dihydroethidium, by Yeasen (1 mentions)
Dcfh da, by Beyotime (1 mentions)
Mtt solution, by Merck Group (1 mentions)
Cell counting kit 8 (cck8), by Transgene (1 mentions)
Human tnf α elisa kit, by Thermo Fisher Scientific (1 mentions)
Mouse elisa kit, by Thermo Fisher Scientific (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay kit, by Promega (1 mentions)
Spectramax m3, by Molecular Devices (1 mentions)
Sandwich elisa kits, by MultiSciences Biotech (1 mentions)
40 protocols using multi mode microplate reader
1
Evaluating NK Cell Cytotoxicity
2
Fluorometric Analysis of Flying Squirrel Extracts
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Aliquots (2 ml) of each methanolic extract were concentrated 10-fold using a nitrogen evaporator (Organomation; Berlin MA, USA) and high purity nitrogen (Praxair Peterborough ON, Canada) and analyzed for fluorescent intensity at different wavelengths using a SpectraMax M3. Multi-Mode Microplate reader (Molecular Devices; San Jose, CA, USA). For each sample, 1 ml was extracted using a 1 ml Eppendorf pipette and placed into a polystyrene cuvette for analysis. Associated curves were obtained using SoftMax Pro at a fixed excitation wavelength of 350 nm, scanning for emissions at 400 nm to 600 nm. These parameters were selected based on previous knowledge of fluorescence in flying squirrels, where all specimens appear to elucidate and absorb wavelengths of at least 395 nm and emit pink (ventral) and blue (dorsal) colours [1 (link)].
3
Esketamine Nanoparticle Release Kinetics
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First, 1 mL of esketamine solutions was prepared at concentrations of 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4.0 μg/mL. Absorbance values were detected at 269 nm using a multimode microplate reader (Molecular Devices) and absorbance concentration curves were plotted. 200 μL of ES NPs-HA were collected and incubated in centrifuge tubes with 1 mL of PBS at 37°C. After collecting a portion of the mixed solution every 24 hours, replace it with a fresh PBS buffer. The absorbance values of the collected mixed solution were measured again using the multimode microplate reader and the daily dose of esketamine released was calculated from the absorbance-concentration curve previously plotted and repeated three times for 21 days. The cumulative release is summed over the daily releases for the corresponding number of days.
4
Quantifying TNF-α Release in Cell Lines
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TNF-α release from RAW 264.7 and HMC3 cells was determined using murine and human TNF-α ELISA kits (Peprotech) according to the manufacturer’s instructions. Briefly, the media (1 mL/well) were collected from the sample after exposure to Mn for the designated periods. ELISA was then performed and the optical density of each well measured. The concentration of secreted TNF-α was determined using a multi-mode microplate reader (Molecular Devices) set to 450 nm, with wavelength correction set at 620 nm.
5
Evaluating LipoCER's Cytotoxic Effects on MC38 Cells
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MC38 cells were treated with LipoCER (10 μmol/L) and vehicle liposomes at 37°C, 5%CO2 for 36 hours. MC38 cells were incubated with 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium (Merck-Calbiochem, Burlington, MA) solution (5 mg/mL) for 2 hours. Then, lysis buffer (1 mmol/L HCL and 10% Triton X-100 in isopropanol) was added, and plates were incubated at room temperature and gently shaken at 70 rpm for 30 minutes to lysate the cells and elute 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium dye. The absorbance was then measured at 590 nm using a multi-mode microplate reader (Molecular Devices, Silicon Valley, CA).
6
BBB Permeability Measurement Using Evans Blue
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The permeability of the BBB was evaluated using extravasation of Evans blue dye (EB, E2129, Sigma-Aldrich; Exk = 620 nm, Emk = 680 nm) as previously reported with some modification [10 (link), 31 (link)]. In brief, 1 h after the intravenous injection of 2% EB (5 mL/kg), the mice were intracardially perfused with heparinized 0.9% saline to remove the intravascular dye. The brains were quickly harvested and separated into left and right hemispheres and weighted, homogenized in normal saline, and centrifuged. Then, 1 ml of supernatant was incubated with 50% trichloroacetic acid for 6 h. Then, the optical density of supernatants was measured at 620 nm using a multimode microplate reader (Molecular Devices, USA). The brain content of EB was expressed as the content of EB dye quantity in the brain quantity (μg/g).
7
MTT Assay for Cell Viability
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Cell viability was determined using the MTT assay according to the manufacturer's instructions (Sigma-Aldrich, St Louis, MO, USA). HepG2 cells were dispensed in 96-well plates at a density of 5 × 104 cells per well and stabilized in a 5% CO2, 37°C incubator for 24 h in DMEM. GS-KG9 was treated at concentrations of 0, 50, 100, 300, 500, 700, or 1,000 µg/mL, and the group not treated with the extract was set as the control group. After 24 h of culture, the medium was removed, and 100 µL of fresh medium was added, followed by stabilization for 30 min in a 5% CO2, 37°C incubator. After adding 50 µL of MTT solution (2 mg) and reacting for 5 h in a 5% CO2, 37°C incubator, the medium was removed, and 150 µL of DMSO was added, and the mixture reacted at room temperature for 20 min to dissolve the insoluble formazan crystals. Absorbance was measured with a multi-mode microplate reader (Molecular Devices, San Jose, CA, USA) at a wavelength of 540 nm.
8
Intracellular ROS Determination in HepG2 Cells
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Intracellular ROS levels were determined according to the manufacturer's instruction (Invitrogen, Carlsbad, CA, USA). Briefly, HepG2 cells were dispensed at 5 × 104 cells per well in 96-well plates and pretreated with GS-KG9 at 0, 25, 50, 100 µg/mL. After incubation for 24 h, FBS-free medium was added to the wells for 30 min. Then 10 µM 2′,7′-Dichlorofluorescein diacetate (DCFH-DA) was added, and the subsequent reaction was carried out in a 37°C incubator for 30 min. After treatment with 30 µM GalN, the cells were washed three times with PBS. Fluorescence was monitored with a multi-mode microplate reader (Molecular Devices) at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. The ROS generation percentage was calculated as follows: (A485/535 of treated cells/A485/535 of untreated cells) ×100.
9
Cell Viability Assay Protocol
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Cell culture protocol was the same as above. The specimens with attached cells were gently rinsed with PBS and metastasized to a new 24-well plate at 1, 3, 5, and 7 days. Then, 500 μL MEM α containing 10% CCK-8 reagent (CCK-8, Dojindo, Kumamoto, Japan) was introduced to each well, and incubated for 2 h. The absorbance was measured at 450 nm using a multi-mode microplate reader (Molecular Devices, San Jose, CA, USA).
10
Quantifying Nuclear HDAC Activity
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