96 well black tissue culture plates

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The 96-well black tissue culture plates are a laboratory equipment designed for cell culture applications. These plates provide a standardized format with 96 individual wells, allowing for the simultaneous culture and analysis of multiple samples. The black plate color is intended to minimize background fluorescence and optimize optical clarity for various imaging and detection techniques.

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Lab products found in correlation

Celltiter glo, by Promega (1 mentions) 2 7 bis 2 carboxyethyl 5 and 6 carboxyfluoresceinacetoxymethyl ester, by Thermo Fisher Scientific (1 mentions) Maxisorp microplate, by Thermo Fisher Scientific (1 mentions) Fibronectin, by Merck Group (1 mentions) Bcecf am, by Thermo Fisher Scientific (1 mentions) Fibrinogen, by Merck Group (1 mentions) Fish skin gelatin, by Merck Group (1 mentions)

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96-well plates (1169 citations) Black 96-well plate (105 citations) 96-well culture plates (78 citations) 96-well tissue culture plates (75 citations) Tissue culture plates (24 citations)

4 protocols using 96 well black tissue culture plates

Cytotoxicity, Cytopathic, and IL-8 Assays

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For cytotoxicity assays, MCF7, L929, or B16Bl6 cells were plated in black 96‐well tissue culture plates (Nunc) at 1,000 cells/well 24 h before stimulation. After 72 h of stimulation, cell supernatant was removed and cell viability was determined with CellTiter‐Glo (Promega).
For the cytopathic effect assay, L929 cells were plated in black 96‐well tissue culture plates (Nunc) at 8 × 10⁴ cells/well in DMEM with 2% FCS. The next day, mIFN‐γ or mAFN‐II was added and cells were incubated overnight. EMC virus (ATCC) was added, and 24 h later, cell viability was determined with CellTiter‐Glo (Promega).
To assay IL‐8 secretion by HUVECs, cells were plated at 2 × 10⁴ cells/well in 96‐well plates the day before stimulation. After 24 h of stimulation, supernatant was collected and IL‐8 concentration was determined via Human IL‐8 ELISA MAX kit (BioLegend).

Huyghe L., Van Parys A., Cauwels A., Van Lint S., De Munter S., Bultinck J., Zabeau L., Hostens J., Goethals A., Vanderroost N., Verhee A., Uzé G., Kley N., Peelman F., Vandekerckhove B., Brouckaert P, & Tavernier J. (2020). Safe eradication of large established tumors using neovasculature‐targeted tumor necrosis factor‐based therapies. EMBO Molecular Medicine, 12(2), e11223.

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Neutrophil Adhesion in Endothelial Cells

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HUVEC were cultured in 96-well black tissue culture plates (Thermo Scientific, UK). Twenty-four hours prior to experimentation, HUVEC were subjected to normoxia or hypoxia in the absence or presence of 10 ng/ml TNF-α. Neutrophil adhesion in response to 20 nM PMA or 100 ng/ml lipopolysaccharide (LPS) were measured as previously described (32 (link)). Briefly, neutrophils were cultured under normoxia or hypoxia for 1 h, labeled with 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluoresceinacetoxymethyl ester (Life Technologies, UK) and then added to wells under normoxia or hypoxia. Fluorescence was measured using a Tecan GENios Spectra FLUOR plate reader (Tecan UK Ltd., UK). Adhesion was calculated by comparing the fluorescence of washed wells to initial fluorescence.

Khawaja A.A., Chong D.L., Sahota J., Mikolasch T.A., Pericleous C., Ripoll V.M., Booth H.L., Khan S., Rodriguez-Justo M., Giles I.P, & Porter J.C. (2020). Identification of a Novel HIF-1α-αMβ2 Integrin-NET Axis in Fibrotic Interstitial Lung Disease. Frontiers in Immunology, 11, 2190.

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Capturing Neutrophil Extracellular Traps

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Plate-bound ICs were prepared by coating wells of 96-well black tissue culture plates (ThermoFischer; ref. 165305) with purified monoclonal Ab 3H9 at 5 μg/mL in PBS overnight at 4°C. The 3H9 mouse monoclonal IgG was grown and purified from culture supernatants by Protein A beads, as described previously (52 (link)). The next day, plates were washed once with PBS and blocked with 2% (w/v) IgG- and protease-free BSA (Jackson ImmunoResearch Labs; ref. 001-000-161) and 10 μg/mL poly-L-Lysine in PBS for 1 h at RT, followed by 2 washes with PBS. NETs, prepared from healthy human neutrophils by incubation with hydroxyapatite and isolated according to previously described procedures (53 (link)), were added to the wells at a concentration of 2 μg/mL protein and 4 μg/mL DNA. Following incubation for 1 h at RT, the plates were washed 3 times with HBSS.

Bendorius M., Neeli I., Wang F., Bonam S.R., Dombi E., Buron N., Borgne-Sanchez A., Poulton J., Radic M, & Muller S. (2018). The Mitochondrion-lysosome Axis in Adaptive and Innate Immunity: Effect of Lupus Regulator Peptide P140 on Mitochondria Autophagy and NETosis. Frontiers in Immunology, 9, 2158.

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Integrin-mediated Neutrophil Adhesion Assay

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Fibronectin (Sigma, UK) (for β₁ integrin engagement) and fibrinogen (Sigma, UK) (for α_Mβ₂ integrin engagement) were immobilized in 96-well black MaxiSorp microplates (Thermo Scientific, UK) and blocked with 2% fish-skin gelatin (Sigma, UK), to reduce non-specific neutrophil binding (Fig. S1). HUVEC were cultured in 96-well black tissue culture plates (Thermo Scientific, UK). HC-neutrophils were labelled with 2.5 μM 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluoresceinacetoxymethyl ester (BCECF-AM) (Life Technologies, UK) for 30 minutes at 37 °C. Stained cells were washed twice in the sodium HEPES buffer by centrifugation at 300 g for 5 minutes and resuspended to 1 × 10⁷ neutrophils/ml. 5 × 10⁵ neutrophils were added to wells containing 20 nM PMA and 200 μg/ml IgG for 30 minutes at 37 °C. Fluorescence was measured using a Tecan GENios Spectra FLUOR plate reader (Tecan UK Ltd., UK). Plates were washed and read again. Adhesion was calculated by comparing the fluorescence of washed wells to initial fluorescence.

Khawaja A.A., Pericleous C., Ripoll V.M., Porter J.C, & Giles I.P. (2019). Autoimmune rheumatic disease IgG has differential effects upon neutrophil integrin activation that is modulated by the endothelium. Scientific Reports, 9, 1283.

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