H1299 (1.5 × 105 cells) and MCF-7 (3.0 × 105 cells) were seeded into a Costar® 24-well plate (Corning, NY, USA) and cultured for 24 h at 37 °C with 5% CO2 to form a confluent cell monolayer. After the cells reached confluence, the medium was removed and PBS buffer was used to wash the cells. The monolayer was then scratched with a 200 µL pipette tip, and the cells were treated with 150 µM EA or MDSA at 37 °C and 5% CO2 for 48 h. Using a Lionheart FX automated microscope (BioTek®, Agilent, Santa Clara, CA, USA), the bright field images of the cells were acquired and analyzed.
Costar 24 well plate
Manufactured by Corning
Sourced in United States
The Costar 24-well plate is a laboratory equipment product designed for cell culture and assay applications. It is a multiwell plate with 24 individual wells, each with a consistent surface area and volume, for performing multiple experiments simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
M per buffer, by Thermo Fisher Scientific (2 mentions)
Cgamp elisa kit, by Cayman Chemical (2 mentions)
Mouse multi plex luminex kit, by Merck Group (2 mentions)
Lionheart fx automated microscope, by Agilent Technologies (1 mentions)
Tween 20, by Merck Group (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Mowiol, by Merck Group (1 mentions)
M2128, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Transwell system, by Corning (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Dispase 2, by Roche (1 mentions)
L glutamine, by Lonza (1 mentions)
Cck 8 solution, by Dojindo Laboratories (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Lightcycler 480 sybr green 1 master mix, by Roche (1 mentions)
High pure rna isolation kit, by Roche (1 mentions)
Jetpei, by Polyplus Transfection (1 mentions)
17 protocols using costar 24 well plate
1
Wound Healing Assay with H1299 and MCF-7 Cells
2
Adipocyte Immunofluorescence Imaging Protocol
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Cells were seeded and differentiated in a Costar 24-well plate (Corning) at 100,000 cells/well on 13 mm cover glass. After the incubation, cells were fixed for 30 minutes with 4% paraformaldehyde at room temperature. The cells were then rinsed three times with PBS and permeabilized with PBS bovine serum albumin 1% (GE Healthcare, Wauwatosa, WI, USA) containing Tween 20 0.5% (Sigma) for 40 minutes. Cells were then incubated for 1 hour at room temperature with primary anti-adipophilin antibody (dilution 1/10, mouse monoclonal, AP125; PROGEN Biotechnik GmbH, Heidelberg, Germany). After having been rinsed three times with PBS bovine serum albumin 1%, cells were then incubated for 1 hour at room temperature in the dark with anti-mouse secondary antibody (coupled to Alexa 568 dilution 1/500) and the lipid probe BODIPY® 493/503 at 20 μg/mL (D3922; Thermo Fisher). Cells were then rinsed three times with PBS and mounted with Mowiol® (Sigma).
3
MTT Assay for Cell Viability
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Cells were seeded and differentiated in a Costar 24-well plate (Corning) at 100,000 cells/well. After incubation, 500 μL of PBS 0.1 M containing MTT (2.5 mg/mL; M2128; Sigma) was added in each well and the cells were incubated for 2 hours in the dark at 37°C in the presence of 5% carbon dioxide. The medium was then removed and cells were lysed in a solution containing 20% sodium dodecyl sulfate and 33% N, N-dimethylformamide (pH 4.7). The optical density was then read at 570 nm.
4
Gingival Fibroblast Response to Porphyromonas gingivalis
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Human gingival fibroblasts, at a seeding density of 5 × 105/well, were cultured in a Costar® 24-well plate (Corning Life Sciences, Corning, NY, USA) in D-MEM medium at 37 °C in an atmosphere of 5% CO2. After the incubation period, fresh medium without antibiotics was added to HGFs, before they were treated with P. gingivalis. HGFs were stimulated with bacteria, at multiplicities of infection (MOI) of 1:100 for 24 h, and with cystatin C at a concentration of 0.3 µg/mL at 37 °C for 24 h, to perform cytokine assays, and evaluate ROS, and NO. Control groups include HGFs without stimulation or stimulated with LPS and peptidoglycans.
5
Epithelial Sheet Isolation for Ex Vivo Culture
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Vaginal and skin tissue was freshly processed for each experiment. To obtain epithelial sheets, vaginal and skin tissue was treated with dispase II (3 U/ml for vaginal tissue and 1 U/ml for skin; Roche Diagnostics), after which the epithelium/epidermis was separated from the rest of the tissue. Ex vivo vaginal epithelial explants were extensively washed, cut to size, and placed in a Transwell system (6.5 mm Transwell, 5.0 μm pore polycarbonate membrane inserts; Corning) complete medium (IMDM; Thermo Fisher Science) supplemented with 10% FCS (Invitrogen), L‐glutamine (2 mM; Lonza), penicillin (10 U/ml; Invitrogen), and streptomycin (10 mg/ml; Invitrogen), containing the according stimuli. Ex vivo skin explants were prepared in a Costar® 24‐well plate (Corning) in complete medium containing the according stimuli as described before (Hertoghs et al, 2019 (link)).
6
Quantifying cGAMP in Transfected HEK 293T Cells
7
Cell Attachment Evaluation on Materials
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To study the attachment of MC3T3-E1 cells on the material surfaces, cell counting kit-8 (CCK-8) assay was performed to analyze the numbers of attached cells on material surfaces cultured for 6, 12, and 24 hours. Before seeding the cells, three disks for each group were put into the wells of a Costar 24-well plate (Corning Incorporated, NY, USA). The cell suspension was seeded into each well at a density of 3×104 viable cells per cm2 of sample, with DMEM as a blank control. The culture plates were incubated at 37°C in a humidified atmosphere of 5% CO2. At each time point, the samples were gently rinsed with phosphate-buffered saline (PBS) to remove the unattached cells then transferred to a new 24-well plate, and 50 μL CCK-8 solution (Dojindo Molecular Technologies Inc., Kumamoto, Japan) was added to each well. The plates were then incubated for 3 hours. At the end of this period, 100 μL of the supernatant was transferred into a 96-well plate. The plates were then read at 450 nm (with 620 nm as the reference wavelength) using a Synergy HT microplate reader (Bio-Tek Instruments Ltd, Winooski, VT, USA). The mean optical density (OD) obtained from the blank control was subtracted from the ODs of the test groups.
8
Allogeneic T Cell Functional Assay
9
Cell Line Growth Rate Measurement
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The growth rate of the stable cell lines expressing WT, K39R, K39Q, K39E Cytc, and EV were measured. Cells were seeded at 30,000 cells/well onto a 0.1% gelatin coated Costar 24-well plate (#3524, Corning Incorporated) and cultured in 2 mL/well growth media. Every 24 h, cells were washed with 1x PBS and collected via trypsinization. Collected cells were counted using a Moxi Z mini automated cell counter (#MXZ001, Orflo Technologies; Ketchum, ID, USA).
10
Quantifying cGAMP in Transfected HEK 293T Cells
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HEK 293T cells were seeded in Costar 24-well plate (Corning, 3526) at 2 × 105 cells per well and cultured at 37 °C with 5% CO2. After 24 h, the cells were transfected with 1.5 ng pcDNA3.1-human cGAS plasmids or 0.05ng pcDNA3.1-mouse cGAS plasmids using the Lipofectamine 2000 according to the manufacture’s manual. Twenty four hours later, the cells were further transfected with 10 μg/ml salmon sperm DNA using Lipofectamine 2000 or with Lipofectamine 2000 alone. After 4 h, the cells were washed with 1×PBS once and lysed in 200 μl M-PER™ buffer (Thermo Fisher Scientific, 78501). After centrifugation at 15000 rpm for 10 mins at 4 °C, cGAMP in the supernatant was quantified using cGAMP ELISA kit (Cayman Chemical, 501700) according to the manufacture’s manual.
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