96 well culture plate

Manufactured by Eppendorf

Sourced in Italy, Germany

The 96-well culture plate is a laboratory equipment designed for cell and tissue culture applications. It consists of a flat, rectangular plate with a grid of 96 individual wells, each capable of holding a small volume of liquid or cell culture media. The plate is made of high-quality, autoclavable materials to ensure sterility and reliability during experiments.

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Lab products found in correlation

Catalase from bovine liver, by Merck Group (1 mentions) Human ifn gamma quantikine elisa, by R&D Systems (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) Thapsigargin, by Santa Cruz Biotechnology (1 mentions) Cck 8, by Beyotime (1 mentions) Glowmax navigator microplate luminometer, by Promega (1 mentions) Polyfect transfection reagent, by Qiagen (1 mentions) Epmotion 5075, by Eppendorf (1 mentions)

Spelling variants of this product

96-well plates (90 citations) 96-well cell culture plates (11 citations) 96-well tissue culture plates (3 citations)

4 protocols using 96 well culture plate

CD4+ T Cell Proliferation and IFNγ Production

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CD4⁺ T cells were labelled with 2 μM CellTrace Violet (Invitrogen, Thermo Fisher Scientific). CD4⁺ T cells and neutrophils were cultured at a 1:1 ratio, 50 000 cells of each cell type, in a 96-well culture plate (Eppendorf) coated with anti-CD3 (clone OKT3, Invitrogen, Thermo Fisher Scientific, 1:500) and anti-CD28 (clone CD28.2, Invitrogen, Thermo Fisher Scientific, 1:1000), in total volume 200 μl RPMI supplemented with 10% fetal calf serum and 2 mM L-glutamine. When indicated, 100 μg/ml catalase (from bovine liver, Sigma) was added to the culture medium. After 3 days of culture at 37°C (5% CO₂) T cell proliferation was analyzed on a Cytoflex flow cytometer (Beckman Coulter).
Levels of IFNγ in culture supernatants were quantified using Human IFN-Gamma Quantikine ELISA (R&D Systems) according to manufacturer’s instructions. The T cell culture supernatants were collected after 3 days of culture and stored at -20°C until analysis.

Arve-Butler S., Mossberg A., Schmidt T., Welinder C., Yan H., Berthold E., Król P, & Kahn R. (2022). Neutrophils Lose the Capacity to Suppress T Cell Proliferation Upon Migration Towards Inflamed Joints in Juvenile Idiopathic Arthritis. Frontiers in Immunology, 12, 795260.

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Synergistic effects of ALA and BTZ

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NB cell lines were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C and 5% CO 2 . ALA was added 24 h before the addition of BTZ (50 nM) for ALA/BTZ combined treatment. For estimation of the effect of BTZ on ER-stress markers and HO-1 expression, NB cells were seeded in six-well culture plate at density 5 × 10 5 cell per well and treated with BTZ alone and in combination with 5 mM 4-sodium phenylbutyrate (4PBA, Sigma-Aldrich, Milan, Italy) for 6 and 24 h, with 10 μM thapsigargin (Santa Cruz Biotechnology) alone and in combination with 5 mM 4-PBA for 24 h. For viability assay, NB cells were seeded on 96-well culture plate (Eppendorf, Milan, Italy) at density 1 × 10 4 cell per well, and subsequently treated with 100 μM of ALA. After 24 h, 50 nM BTZ alone and in combination with ALA was added to cell cultures for 24 h. All agents were diluted directly in cell culture medium.

Tibullo D., Giallongo C., Puglisi F., Tomassoni D., Camiolo G., Cristaldi M., Brundo M.V., Anfuso C.D., Lupo G., Stampone T., Li Volti G., Amenta F., Avola R, & Bramanti V. (2018). Effect of Lipoic Acid on the Biochemical Mechanisms of Resistance to Bortezomib in SH-SY5Y Neuroblastoma Cells. Molecular neurobiology, 55(4).

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NaF Impacts on PDLCs Proliferation

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Effects of various concentrations of NaF (Sigma, USA) on PDLCs proliferation was measured using a cell counting kit (CCK-8; Beyotime, China). PDLCs were seeded into 96-well culture plates (Eppendorf) with a concentration of 1×10³ cells/well. After 24-h incubation at 37°C with 5% CO₂, the plates were treated with 0, 1, 5, 10, 50, 100, 5×10², 1×10³, and 5×10³ μmol/L NaF for 1, 2, 3, 4, 5, and 6 days. CCK-8 was mixed with serum-free ɑ-MEM medium at a proportion of 1:10 in advance. After removal of complete ɑ-MEM medium, 110 μL mixture was added to each well and incubated at 37°C for 2 h, until the media turned yellow. Groups without cells were used as zero setting. We assessed cell viability by absorbance values in each well, which was measured with a spectrophotometer (Thermo, Finland) at a wavelength of 450 nm. Data were calculated using averages of three wells, and untreated PDLCs were considered as the control group. The concentrations of 0, 10, 5×10² and 1×10³ μmol/L were chosen for subsequent experiments.

Li K.Q., Jia S.S., Ma M., Shen H.Z., Xu L., Liu G.P., Huang S.Y, & Zhang D.S. (2016). Effects of fluoride on proliferation and mineralization in periodontal ligament cells in vitro. Brazilian Journal of Medical and Biological Research, 49(8), e5291.

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Automated Dual-Luciferase Reporter Assay

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To enhance the reproducibility and replicability of results, we utilized the automatic liquid handling system epMotion 5075 (Eppendorf, Hamburg, Germany) for the Dual-Luciferase Reporter Assay in this study. Specifically, 3.6 × 10⁴ HEK-293T cells were seeded per well of the 96-well culture plates (Eppendorf) and were transfected 24 h later with 1 µL/well PolyFect transfection reagent (Qiagen, Hilden, Germany) containing 50 ng/well of each reporter vector and 200 ng/well of each expression vector. For each run, empty vector constructs (pMIR-empty and pSG5-empty) were included as a control, and all plasmid combinations were transfected in technical duplicates. After 48 h, cells were lysed and measured using the Dual-Luciferase Reporter Assay System manual with the GlowMax navigator microplate luminometer (Promega, Madison, WS, USA). We performed Luciferase assays of the WT 3’UTRs in four independent experiments and compared WT to mutated binding sites within the 3′UTR in three independent experiments. As a positive control, we re-evaluated the downregulation of four target genes (PFKFB4, HMMR, UBQLN3, and ODF2) previously studied by overexpressing microRNA-23a/b-3p using this automated technique, as shown in Figure S1.

Becker L.S., Al Smadi M.A., Koch H., Abdul-Khaliq H., Meese E, & Abu-Halima M. (2023). Towards a More Comprehensive Picture of the MicroRNA-23a/b-3p Impact on Impaired Male Fertility. Biology, 12(6), 800.

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