96 multiwell black plate

Manufactured by Corning

Sourced in United States

The 96 multiwell black plate is a laboratory equipment used for various assays and experiments. It has 96 individual wells arranged in a 8x12 grid format. The plate is made of black-colored material, which helps reduce background fluorescence and luminescence, enhancing signal detection. The wells are designed to accommodate small sample volumes, making it suitable for high-throughput screening and microplate-based applications.

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Lab products found in correlation

Glomax multi detection system plate reader, by Promega (2 mentions) Spectramax m5 microplate reader, by Molecular Devices (1 mentions) Celltiter 96 aqueous, by Promega (1 mentions) X pert pro, by Philips (1 mentions) Opteia, by BD (1 mentions) Caspase glo 3 7 assay kit, by Promega (1 mentions) Fluo 4 nw calcium assay kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

96-multiwell plate (9 citations) Black wall/clear bottom 96-multiwell plates (5 citations)

7 protocols using 96 multiwell black plate

Cell Viability Assay of Nanoparticles

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Cell viability
was determined
by an MTS assay, which was carried out with CellTiter 96 AQueous (Promega
Corp.) kit.^{25 (link)} There were 1 × 10⁴ BEAS-2B or RAW 264.7 cells in 100 μL of culture medium
plated in each well of a 96 multiwell black plate (Costar, Corning,
NY) for overnight growth. The medium was removed, and cells were treated
for 24 h with 100 μL of 50, 100 and 200 μg/mL nanoparticle
suspensions. After the treatment, the cell culture medium was removed
and followed by washing of the plates three times with PBS. Each well
received 100 μL of culture medium containing 16.7% of MTS stock
solution for 1 h at 37 °C in a humidified 5% CO₂ incubator.
The plate was centrifuged at 2000g for 10 min in
NI Eppendorf 5430 with a microplate rotor to spin down the cell debris.
A 80 μL amount of the supernatant was removed from each well
and transferred into a new 96 multiwell plate. The absorbance of formazan
was read at 490 nm on a SpectraMax M5 microplate reader (Molecular
Devices Corp., Sunnyvale, CA, USA).

Zhang H., Pokhrel S., Ji Z., Meng H., Wang X., Lin S., Chang C.H., Li L., Li R., Sun B., Wang M., Liao Y.P., Liu R., Xia T., Mädler L, & Nel A.E. (2014). PdO Doping Tunes Band-Gap Energy Levels as Well as Oxidative Stress Responses to a Co3O4p-Type Semiconductor in Cells and the Lung. Journal of the American Chemical Society, 136(17), 6406-6420.

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Graphene Oxide Oxidation and Reactive Species

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The oxidation capacity and oxygen species of GO samples were determined as described before^{25 (link)}. 2′,7′-dichlorodihydro- uorescein diacetate (H₂DCF-DA) was used to evaluate the total oxidation capacity of GO, while the 3′-(p-aminophenyl) fluorescein (APF) and 3-bis(2-methoxy-4-nitro-5-sulfophehyl)-2H-tetrazolium-5-carboxanilide) (XTT) assays were used to determine OH and O₂⁻ on GO surface, respectively. The DCF working solution was prepared by mixing 50 μg of H₂DCF-DA with 280 μL 0.01 M NaOH. The resulting solution was incubated for 30 min, and diluted with 1720 μL of a sodium phosphate buffer (25 mmol/L, pH = 7.4) to form 25 μg/mL DCF solution. 95 μL aliquots of 25 μg/mL DCF, 100 μM XTT or 10 μM APF working solutions were added into each well of a 96 multiwell black plate (Costar, Corning, NY). A 5 μL amount of 5 mg/mL nanoparticle suspension was subsequently added to each well, followed by 2 h incubation. DCF uorescence emission spectra in the range of 500–600 nm were collected using a SpectraMax M5 microplate reader with an excitation wavelength of 490 nm. APF uorescence emission spectra were collected at 480–600 nm with an excitation wavelength of 455 nm, while XTT absorbance spectra were recorded in the range of 410–550 nm.

Li R., Mansukhani N.D., Guiney L.M., Ji Z., Zhao Y., Chang C.H., French C.T., Miller J.F., Hersam M.C., Nel A.E, & Xia T. (2016). Identification and Optimization of Carbon Radicals on Hydrated Graphene Oxide for Ubiquitous Antibacterial Coatings. ACS nano, 10(12), 10966-10980.

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Characterization of Fe2O3 Nanoparticles

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A particle suspension (50 μg/mL in DI H₂O) drop was added on the TEM grids for air-dry at room temperature. The TEM observation was performed on a JEOL 1200 EX instrument (accelerating voltage 80 kV). A Philips X’Pert Pro diffractometer equipped with CuKr radiation were used to obtain the XRD spectra. The hydrodynamic diameter and surface charge in water and cell culture media were measured by DLS coupled with ZPA (Brookhaven Instruments Corporation, Holtsville, NY, USA). DCF assay was used to evaluate the surface reactivity of Fe₂O₃ particles. In detail, 50 μg of H₂DCF-DA were mixed with 280 μL 0.01 M NaOH and incubated for 30 min at room temperature. The resulting solution was diluted with 1720 μL of a sodium phosphate buffer (25 mmol/L, pH = 7.4) to form 25 μg/mL DCF working solution. A 5 μL aliquots of nanoparticle suspension (5 mg/mL) were added into each well of a 96-multiwell black plate (Costar, Corning, NY) and then 95 μL amount of DCF working solution was added to each well, followed by 2 h incubation. DCF fluorescence emission spectra were recorded by a SpectraMax M5 microplate reader at an excitation wavelength of 490 nm^{8 (link)}.

Cai X., Dong J., Liu J., Zheng H., Kaweeteerawat C., Wang F., Ji Z, & Li R. (2018). Multi-hierarchical profiling the structure-activity relationships of engineered nanomaterials at nano-bio interfaces. Nature Communications, 9, 4416.

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Evaluating Oxidative Capacity of MOx

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The oxidative capacity of MOx were determined by a method as described before [15 (link)]. 2’,7’-dichlorodihydro-fluorescein diacetate (H₂DCFDA) was used to evaluate the oxidation capacity of MOx. The 2',7'-dichlorofluorescein (DCF) working solution was prepared by mixing 50 μg of H₂DCFDA with 280 μL 0.01 M NaOH. The resulting solution was incubated for 30 min, and diluted with 1720 μL of a sodium phosphate buffer (25 mmol/L, pH = 7.4) to form 25 μg/mL DCF solution. 96 μL aliquots of 25 μg/mL DCF working solutions were added into each well of a 96 multiwell black plate (Costar, Corning, NY). A 4 μL amount of 5 mg/mL nanoparticle suspension was subsequently added to each well, followed by 2 h incubation. DCF fluorescence emission spectra in the range of 500–600 nm were collected using a SpectraMax M5 microplate reader with an excitation wavelength of 490 nm.

Cai X., Lee A., Ji Z., Huang C., Chang C.H., Wang X., Liao Y.P., Xia T, & Li R. (2017). Reduction of pulmonary toxicity of metal oxide nanoparticles by phosphonate-based surface passivation. Particle and Fibre Toxicology, 14, 13.

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Nanoparticle-Induced Cytokine Release Assay

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There
were 1 × 10⁴ BEAS-2B
or RAW 264.7 cells in 100 μL culture medium plated overnight
in each well of a 96 multiwell black plate (Costar, Corning, NY).
The medium was removed, and cells were treated with each of the nanoparticle
suspensions at 50, 100, and 200 μg/mL for 6 h. Plates were centrifuged
at 2000g for 10 min in an Eppendorf 5430 microcentrifuge
with microplate rotor to spin down the cell debris and nanoparticles.
A 50 μL amount of the supernatant was removed from each well
for measurement of IL-8 activity in BEAS-2B cells or TNF-α
activity in RAW 264.7 cells using an OptEIA (BD Biosciences, CA)
ELISA kit according to the manufacturer’s instructions. Briefly,
a 96-well plate was coated with 50 μL of monoclonal anti-IL-8
or anti-TNF-α antibody for 2 h. After removal of the unbound
antibody, a standard cytokine dilution series or 50 μL of each
supernatant were pipetted into the precoated wells for antigen capture.
After 2 h, the unbound growth factor was removed and each well in
the plate was washed with a buffer five times and an enzyme-linked
secondary polyclonal antibody added. Following washing, a substrate
solution was added at 1:250 dilution for 30 min to allow color development.
After termination of the reaction, the colorimetric intensity was
measured at 450 nm on a SpectraMax M5 microplate reader.

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Caspase 3/7 Activity Quantification

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Caspase 3/7 activity was carried out using the Caspase-Glo 3/7 assay kit (Promega, UK) according to the manufacturer’s protocol. Briefly, BMDM at 3 × 10⁴ cells per well in a black 96-multiwell plate (Corning, NY, USA) were incubated with different stimuli for 6 h. Then, 100 µl of Caspase-Glo 3/7 reagent was added to the wells. Plates were gently shaken and then incubated in the dark at 20°C for 60 min before luciferase activity was recorded using a GloMax^®-Multi Detection System plate reader (Promega, UK).

Garrido D., Chanteloup N.K., Trotereau A., Lion A., Bailleul G., Esnault E., Trapp S., Quéré P., Schouler C, & Guabiraba R. (2017). Characterization of the Phospholipid Platelet-Activating Factor As a Mediator of Inflammation in Chickens. Frontiers in Veterinary Science, 4, 226.

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Fluo-4 NW Calcium Assay in HD11 Cells

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The Fluo-4 NW Calcium Assay kit (Molecular Probes, USA) was used to measure changes in HD11 cells’ intracellular calcium levels upon different stimuli. Briefly, HD11 cells at 3 × 10⁴ cells per well in a black 96-multiwell plate (Corning, NY, USA) were loaded with 100 µl of the dye loading solution containing Fluo-4 NW dye and probenecid, according to the manufacturer’s instructions. The 96-well plate was incubated at 40°C under 5% CO₂ for 30 min in the dark, and the stimuli of interest were added to the cells at T = 0, and the plates were read immediately. The changes in Fluo-4 NW fluorescence were measured at an excitation wavelength of 494 nm and an emission wavelength of 516 nm in a GloMax^®-Multi Detection System plate reader (Promega, UK). Calcium mobilization was recorded over time (213 s, with 3 s intervals).

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