96 well black wall plates

Manufactured by Corning

Sourced in United States

The 96-well black-wall plates are a type of laboratory equipment used for various assays and experiments. These plates feature a black-colored wall, which helps to minimize light interference and background fluorescence, improving the sensitivity and accuracy of the measurements. The plates are designed with 96 individual wells, allowing for multiple samples or replicates to be tested simultaneously. The core function of these plates is to provide a standardized and controlled environment for conducting various biological, biochemical, or analytical procedures.

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Lab products found in correlation

Infinite m1000 pro, by Tecan (2 mentions) Britelite plus luciferase reagent, by PerkinElmer (2 mentions) Luminometer, by PerkinElmer (2 mentions) 2 7 dichlorofluorescein diacetate dcf da, by Merck Group (1 mentions) Spectramax i3, by Molecular Devices (1 mentions) Synergy 2 multi mode microplate reader, by Agilent Technologies (1 mentions) Basic fibroblast growth factor (bfgf), by Roche (1 mentions) Lipid supplement, by Thermo Fisher Scientific (1 mentions) E plate, by Agilent Technologies (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions)

7 protocols using 96 well black wall plates

Oxidative Stress Induction in MB49 Cells

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MB49 cells were seeded in 96-well black wall plates
(Corning, NY, USA) at a concentration of 1 × 10⁵ cells/well
and exposed to a complete medium with Ti(OH)₄ activated
for 1 h with visible light. A solution of 10 μmol/L of H₂O₂ was used (30 min of exposure) as a positive
control to induce oxidative stress in the cells. Cells without any
treatment were used as a negative control. After the exposure period,
the cells were washed with PBS (1×) and labeled with 30 μL
of a 100 μmol/L solution of 2′,7′-dichlorofluorescein
diacetate (DCF-DA) (Sigma-Aldrich, USA). Fluorescence readings were
taken using the Spectra Max i3 (Molecular Devices) with 485–530
nm excitation.^{74 (link)}

Robeldo T., Ribeiro L.S., Manrique L., Kubo A.M., Longo E., Camargo E.R, & Borra R.C. (2022). Modified Titanium Dioxide as a Potential Visible-Light-Activated Photosensitizer for Bladder Cancer Treatment. ACS Omega, 7(21), 17563-17574.

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Quantifying Intracellular Photosensitizer Levels

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Fresh PECA cells (1 × 10⁵ cells/well) were seeded in 96-well black wall plates (Corning Inc, Corning, NY, USA) and cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 IU mL^{− 1}), streptomycin (100 μg mL^{− 1}) for 24 h. Attached cells were washed twice with PBS and then incubated with ALA of different concentrations (0.1 to 10 mM) in 100-μL serum-free medium in the dark with different incubation times (1 to 24 h). PpIX fluorescence emission from the cells was detected using a microplate reader (Synergy 2 multi-mode microplate reader; BioTek, Winooski, VT, USA). The fluorescence excitation wavelength was set at 400 nm and the emission wavelength at 635 nm. Data were corrected for autofluorescence background (n = 5).

Ji J., Fan Z., Zhou F., Wang X., Shi L., Zhang H., Wang P., Yang D., Zhang L., Chen W.R, & Wang X. (2015). Improvement of DC vaccine with ALA-PDT induced immunogenic apoptotic cells for skin squamous cell carcinoma. Oncotarget, 6(19), 17135-17146.

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HEK293T Cell Fluorescence Assay

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10,000 HEK293T cells per well were seeded in 96-well black-wall plates (Corning). 16–24 hours post-seeding, cells were transfected at approximately 60% confluency with .5 μL of Lipofectamine 2000 according to the manufacturer’s protocols and 100 ng of dCas9–VPR plasmid, 30 ng of GFP reporter plasmid, 15 ng of iRFP plasmid, and 20 ng of sgRNA, pegRNA, or epegRNA plasmid. After three days, cells were measured for GFP and iRFP fluorescence using an Infinite M1000 Pro microplate reader (Tecan). GFP fluorescence was normalized to iRFP fluorescence after subtracting background fluorescence signal from untreated cells.

Nelson J.W., Randolph P.B., Shen S.P., Everette K.A., Chen P.J., Anzalone A.V., An M., Newby G.A., Chen J.C., Hsu A, & Liu D.R. (2021). Engineered pegRNAs improve prime editing efficiency. Nature biotechnology, 40(3), 402-410.

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HEK293T Cell Fluorescence Assay

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Culturing hCMEC/D3 Brain Endothelial Cells

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The hCMEC/D3 human brain microvascular endothelial cell line was obtained from Merck Millipore (Germany). To maintain the cells’ brain endothelial-like features, we used cells under passage number 35 [16 (link)]. Cells were grown in dishes coated with rat tail collagen and maintained in an incubator at 37°C with 5% CO₂. The basal medium used was MCDB 131 (Pan Biotech, Germany) supplemented with 5% fetal bovine serum, GlutaMAX (100 ×, Life Technologies, USA), lipid supplement (100 ×, Life Technologies, USA), 10 μg/ml ascorbic acid, 550 nM hydrocortisone, 37.5 μg/ml heparin, 1 ng/ml basic fibroblast growth factor (Roche, USA), 5 μg/ml insulin, 5 μg/ml transferrin, 5 ng/ml selenium supplement (100x, PanBiotech), 10 mM HEPES, and gentamycin (50 μg/ml). We changed the medium every two or three days. When the cultures reached almost 90% confluence, we passaged them to rat tail collagen-coated 96-well plates (E-plate, Agilent, USA) for impedance measurement assays, cell culture inserts (0.4 μm pore size, cellQUART, Sabeu, Germany) for barrier integrity assays and 96-well black-wall plates (Corning, USA) for ROS measurement. Before each experiment, the medium was supplemented with 10 mM LiCl for 24 hours to improve BBB properties [17 (link)]. For further characterization and gene expression studies please see [15 (link)–17 (link), 24 (link)].

Vágvölgyi M., Laczkó D., Santa-Maria A.R., Vigh J.P., Walter F.R., Berkecz R., Deli M.A., Tóth G, & Hunyadi A. (2024). 17-Oxime ethers of oxidized ecdysteroid derivatives modulate oxidative stress in human brain endothelial cells and dose-dependently might protect or damage the blood-brain barrier. PLOS ONE, 19(2), e0290526.

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Pseudovirion Infectivity Assay Protocol

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Supernatant was collected from each well and centrifuged for five minutes at 300 x g to remove cells and debris. Virions were harvested by centrifugation of culture supernatants through a 20% sucrose cushion at 23,500 x g for one hour at 4°C. The pelleted pseudovirions were resuspended in 600 μl of complete media and dilutions (1:3 and 1:9) were made for each pseudovirion-preparation. The diluted virions were mixed with 1.25 × 10⁴ TZM-bl cells in 100 μl DMEM per well in 96-well black-wall plates (Corning), in triplicate. The cells were incubated for 48 hours at 37°C. The supernatant was then removed, and cells lysed in 70 μl of 0.5% Triton-X, before addition of 70 μl of Britelite Plus Luciferase reagent (Perkin Elmer). Luminescence was quantified using a Perkin Elmer luminometer. Data are presented as relative light units (RLUs) following subtraction of background controls. Where indicated, data are expressed as RLUs normalized to the concentration of HIV capsid antigen (p24), as measured by ELISA (ABL, Rockville, MD), following the manufacturer’s instructions.

Angerstein A.O., Stoneham C.A., Ramirez P.W., Guatelli J.C, & Vollbrecht T. (2020). Sensitivity to Monoclonal Antibody 447–52D and an Open Env Trimer Conformation Correlate Poorly with Inhibition of HIV-1 Infectivity by SERINC5. Virology, 548, 73-81.

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Pseudovirion Infection Assay Protocol

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Supernatant was collected from each well and centrifuged for five minutes at 300 x g to remove cells and debris. Virions were harvested by centrifugation of culture supernatants through a 20% sucrose cushion at 23,500 x g for one hour at 4°C. The pelleted pseudovirions were resuspended in 600 μl of complete media and dilutions (1:3 and 1:9) were made for each pseudovirion-preparation. The diluted virions were mixed with 2 x 10 4 TZM-bl cells in 100 μl DMEM per well in 96-well black-wall plates (Corning), in triplicate.
The cells were incubated for 48 hours at 37°C. The supernatant was then removed, and cells lysed in 70 μl of 0.5% Triton-X, before addition of 70 μl of Britelite Plus Luciferase reagent (Perkin Elmer).
Luminescence was quantified using a Perkin Elmer luminometer. Data are presented as relative light units (RLUs) following subtraction of background controls.

Angerstein A.O., Stoneham C.A., Ramirez P.W., Guatelli J., & Vollbrecht T. (2020). Sensitivity to Monoclonal Antibody 447-52D and an Open Env Trimer Conformation Correlate Poorly with Inhibition of HIV-1 Infectivity by SERINC5.

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