96 well cell culture cluster

Manufactured by Corning

Sourced in United States

The 96-well cell culture cluster is a laboratory equipment designed for culturing cells in a high-throughput manner. It provides a standardized 96-well format for efficient cell growth and experimentation. The product features a durable, flat-bottom design with a total well volume of 300-400 μL, suitable for a variety of cell types and applications.

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Lab products found in correlation

Cell counting kit 8 (cck8), by Dojindo Laboratories (3 mentions) Cck 8 reagent, by Dojindo Laboratories (2 mentions) Digital camera, by Nikon (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Corning (1 mentions) Multiskan mk3, by Thermo Fisher Scientific (1 mentions) Fluorescent microscope, by Olympus (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by RiboBio (1 mentions) Multiskan go, by Thermo Fisher Scientific (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Infinite m1000 pro, by Tecan (1 mentions) Rwnt5a, by R&D Systems (1 mentions) Kaempferol, by Merck Group (1 mentions) Powershot g10 camera, by Canon (1 mentions) Azd9496, by Selleck Chemicals (1 mentions) Herceptin, by Genentech (1 mentions)

Spelling variants of this product

96 wells (10 citations) 96-well cell culture cluster plates (6 citations) 96-Well Cell Culture Cluster Costar 3599 (2 citations)

14 protocols using 96 well cell culture cluster

Pseudovirus Infection Assay with MSCs

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Human MSCs were incubated with pseudovirus at a density of 1 × 10⁴ using 96 well cell culture cluster (Costar, 3599). After 48 h, cells were harvested for luciferase experiments (Vigorous, Dual-Lucy Assay Kit).
Different cell lines including 293 T, NCM460 and Beas-2B cells (lower chamber) were co-cultured with MSCs (upper chamber) at the density of 2 × 10⁵ using Transwell BD Matrigel (Costar, 3450). After 3 days, cells were incubated with pseudovirus at the density of 1 × 10⁴ using 96 well cell culture cluster (Costar, 3599). After 48 h, cells were harvested for luciferase experiments (Vigorous, Dual-Lucy Assay Kit).

Cao Y., Wu H., Zhai W., Wang Y., Li M., Li M., Yang L., Tian Y., Song Y., Li J., Wang Y., Ding Q., Zhang L., Cai M, & Chang Z. (2020). A safety consideration of mesenchymal stem cell therapy on COVID-19. Stem Cell Research, 49, 102066.

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Hemolytic Assay for Antimicrobial Peptides

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The methodology used for hemolytic assay was based on Onuma et al. [37 (link)]. Fresh human red blood cells (RBC) (O +) were washed three times with saline buffer (2.76 g/L de NaH₂PO₄ and 9 g/L NaCl, pH 7.4). A suspension of erythrocytes 1% v/v and peptides (at 25, 50, 75 and 100 mM) were prepared with the Tris-saline buffer. Aliquots of 100 μL blood suspension and 100 μL peptide solution were incubated at 37°C for 1 hour. The samples were centrifuged at 3000 x g for 2 min. Aliquots of 100 μL were added to the wells of “96-well cell culture cluster—Costar^®3599” and the absorbance was determined at 405nm in a microplate reader model 3550 Bio-Rad^®_. As a positive control for hemolysis (100% lysis), 1% (v/v) Triton X-100 solution was used. PBS buffer was used as a negative control (0% lysis). The assay was performed in triplicate to determine the percentage of hemolysis at different concentrations of peptides. High concentrations of peptide were used for all cytotoxicity assays with the objective to verify the possible effect of these AMP against mammalian cells in extreme situation. Hemolysis percentage was calculated as follows:

% H e m o l y s i s = [\frac{(A b s 405 n m o f A M P S o l u t i o n - A b s 405 n m o f P B S)}{(A b s 405 n m T r i t o n X 100 - A b s 405 n m o f P B S)}] x 100

Inui Kishi R.N., Stach-Machado D., Singulani J.D., dos Santos C.T., Fusco-Almeida A.M., Cilli E.M., Freitas-Astúa J., Picchi S.C, & Machado M.A. (2018). Evaluation of cytotoxicity features of antimicrobial peptides with potential to control bacterial diseases of citrus. PLoS ONE, 13(9), e0203451.

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Effect of gEGF on Chicken Fibroblast Proliferation

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The UMNSAH/DF-1 cells (ATCC-CRL-12203), a chicken embryo fibroblast line (BeNa Culture Collection Co., Ltd., Beijing, China), were cultured for 24 h in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA) until they attained a full monolayer and then diluted at 1:10. Next, 100 μL of the diluted cell culture was added into each well of a 96-well Cell Culture Cluster (Corning Inc., Shanghai, China) and cultured for 12 h in DMEM at 37 °C to achieve 60% of the cells of the full monolayer and then starved for 12 h in DMEM without fetal bovine serum at 37 °C. The cell cultures were washed with 100 μL PBS before being treated with 100 μL DMEM containing gEGF (1, 5, 10, and 20 μg/mL) for 12 h, while the control was treated with 100 μL DMEM without gEGF. Finally, 10 μL Cell Counting Kit-8 (Dojindo Molecular Technologies, Inc., Rockville, MD, USA) was added to the cell cultures and incubated for 1 h. The OD₄₅₀ of the cultures was measured using a MULTISKAN MK3 (Thermo Electron Corporation, Waltham, MA, USA).

Zhou Y., Chen P., Shi S., Li X., Shi D., Zhou Z., Li Z, & Xiao Y. (2021). Expression of Gallus Epidermal Growth Factor (gEGF) with Food-Grade Lactococcus lactis Expression System and Its Biological Effects on Broiler Chickens. Biomolecules, 11(1), 103.

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Cell Proliferation Assay Protocol

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Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay following the manufacturer's instructions (Dojindo Molecular Technologies, Inc., Kumamoto, Japan). In brief, cells that were transfected with small interfering RNA (siRNA) were seeded at a density of 2×10⁴/well into a 96-well cell culture cluster (Corning Incorporated, Corning, NY, USA) in 100 µl culture medium and incubated overnight. CCK-8 reagents were added to a subset of the wells, and they were incubated for 2 h at 37°C. The absorbance was recorded with an automated plate reader. Each experiment was performed in triplicate and repeated a minimum of three times.

ZHANG Z., MENG G., WANG L., MA Y, & GUAN Z. (2016). The prognostic role and reduced expression of FOXJ2 in human hepatocellular carcinoma. Molecular Medicine Reports, 14(1), 254-262.

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Taxol Cytotoxicity Assay Protocol

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Cells (5×10⁴/ml) sub-cultured in a 96-well cell culture cluster (Corning, Tewksbury, MA, USA) were treated with different concentrations of taxol. MTT (5 mg/ml, 20 μl) was added to each well 4 h prior to the indicated time points. Following 4 h of incubation at 37°C, the medium was removed and the precipitate was dissolved in dimethylsulfoxide. Then, the optical density (OD) values were measured at 570 nm using an ELISA reader (Multiskan MK3, Shanghai Bio-excellent, Shanghai, China). Relative cell viability was calculated according to the following formula: Cell relative viability (%) = OD^experiment/OD^control × 100% (OD blank was used to zero). The IC₅₀ was defined as the drug concentration required to decrease the cell viability to 50% of the control (no drug) value.

LU S., YU L., MU Y., MA J., TIAN J., XU W, & WANG H. (2014). Role and mechanism of Twist1 in modulating the chemosensitivity of FaDu cells. Molecular Medicine Reports, 10(1), 53-60.

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Cell Proliferation Assays: EdU Incorporation and CCK-8

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MDA-MB-231 cells grown on 96-well plate were cultured in triplicate in 96-well plates at a density of 1×10³ for 12 h. Then, cells were labeled with 30 µM 5-ethynyl-2′-deoxyuridine (Ribobio, Guangzhou, China) for 1 h at 37°C. The cells were fixed with 50 µl 4% formaldehyde for 30 min and 0.5% Triton X-100 for 20 min for permeabilization. One hundred microliters 1× Apollo reaction cocktail was added to each well for 30 min after washing with PBS thrice. Then, cells were stained with 100 µl Hoechst 33342 for 30 min and washed with PBS. The stained cells were visualized under a fluorescent microscope (Olympus Corporation, Tokyo, Japan) and counted with Photoshop (Adobe Systems, Inc., San Jose, CA, USA). The incorporation rate represents EdU fluorescence against DNA content.
(2 (link)). Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan) was employed to assess cell proliferation. In brief, cells were seeded on a 96-well cell culture cluster (Corning Inc., Corning, NY, USA) at a concentration of 2×10⁴/well in a volume of 100 µl, and grown overnight. Cell Counting Kit-8 (Dojindo) reagents were then added, incubated for 2 h at 37°C, and absorbance was quantified on an automated plate reader. Each experiment was performed in triplicate and repeated at least three times.

Sheng C., Qiu J., Wang Y., He Z., Wang H., Wang Q., Huang Y., Zhu L., Shi F., Chen Y., Xiong S., Xu Z, & Ni Q. (2018). Knockdown of Ran GTPase expression inhibits the proliferation and migration of breast cancer cells. Molecular Medicine Reports, 18(1), 157-168.

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Cell Viability Assay with Sodium Valproate

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Cells (1.0 × 10⁵/mL) were subcultured in a 96-well cell culture cluster (Corning Inc., Corning, NY, USA) (100μL/well), and each sodium valproate concentration group was set to four replicates. CCK-8 (10 μL) was added into each well 1-4 h before the indicated time points. After 1-4 h of incubation at 37şC, the optical density (OD) values were measured at 535 nm using an ELISA reader (Multiskan GO, Thermo Scientific, MA, USA). Relative cell viability was calculated according to the following formula:

Ma X.J., Wang Y.S., Gu W.P, & Zhao X. (2017). The role and possible molecular mechanism of valproic acid in the growth of MCF-7 breast cancer cells. Croatian Medical Journal, 58(5), 349-357.

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Cell Proliferation Assay for ACHN and A498 Cells

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Proliferation of ACHN and A498 cells was detected using a CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega Corporation) in accordance with the manufacturer's protocol. Briefly, cells were seeded in a 96-well cell culture cluster (Corning Incorporated, Corning, NY, USA) at a density of 3,000 cells per well with 100 µl culture medium. After 5 days, the culture medium was removed and replaced with an equal volume of medium containing CellTiter 96 AQueous One Solution reagent, and the cells were then incubated at 4°C for 2 h. The absorbance was detected at 490 nm using a 96-well plate reader.

Qiu M., Liang Z., Chen L., Tan G., Liu L., Wang K., Chen H, & Liu J. (2017). MicroRNA-200c suppresses cell growth and metastasis by targeting Bmi-1 and E2F3 in renal cancer cells. Experimental and Therapeutic Medicine, 13(4), 1329-1336.

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MTT Assay for Cell Viability Assessment

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HepG2 and SH-SY5Y cell viability was assessed as previously reported by using a conventional 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay [3 (link),34 (link)]. In brief, viable cells were seeded and grown for 24 h into a sterile 96-well cell culture cluster (Corning, Sigma-Aldrich, St. Louis, MO, USA) in a complete medium and maintained at 37 °C in a humidified atmosphere with 5% CO₂. The cells were then added with different concentrations of each compound for a period of 1 h or 24 h. At the end, the culture medium was replaced by a solution of 0.5 mg/mL MTT (Sigma-Aldrich, St. Louis, MO, USA) in PBS. After 2 h of incubation at 37 °C in 5% CO₂, the supernatant was carefully removed from each well, and the formazan crystals were dissolved in DMSO. The absorbance values were measured at 570 nm using Tecan Infinite M1000 Pro (Tecan, Cernusco S.N., Italy) and DMSO medium as blank solution. The results were reported as % cell viability = [optical density (OD) of tested compound/medium OD of control cells] × 100. The results are expressed as mean ± SEM of at least three independent measurements in triplicate.

Carocci A., Barbarossa A., Leuci R., Carrieri A., Brunetti L., Laghezza A., Catto M., Limongelli F., Chaves S., Tortorella P., Altomare C.D., Santos M.A., Loiodice F, & Piemontese L. (2022). Novel Phenothiazine/Donepezil-like Hybrids Endowed with Antioxidant Activity for a Multi-Target Approach to the Therapy of Alzheimer’s Disease. Antioxidants, 11(9), 1631.

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Cytotoxicity Assay for Multiple Myeloma Cells

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For drug cytotoxicity assays, MM cells were washed once and adhered to FN or stromal cells, as previously described [39 (link)]. In our preliminary experiment [39 (link)], the proper drug concentration needed to induce apoptosis in our cell lines had already been established. The viability of MM cells following addition of Doxorubicin, Mitoxantrone or Dexamethasone, at varying concentrations (1 μM, 2 μM and 0.05 μM), for 72 h, was assessed using Cell Counting Kit-8 (CCK8) assay and trypan staining (*, P < 0.05 compared with the group of control). For drug exposure, after 24 h, drugs or vehicle control were added to each well and incubated for an additional 72 h, after which the medium containing drugs was removed and the suspended and attached MM cells were collected.
To evaluate cell viability, cells were seeded on a 96-well cell culture cluster (Corning Inc., Corning, NY) at a concentration of 5 × 10⁴/well in a volume of l L and were grown overnight. CCK8 reagents (Dojindo, Kumamoto, Japan) were added to the different subset wells and the cells were incubated at 37 °C and 5% CO₂. The absorbance was quantified using an automated plate reader at a test wavelength of 450 nm for at least three times.

Huang Y., Huang X., Cheng C., Xu X., Liu H., Yang X., Yao L., Ding Z., Tang J., He S, & Wang Y. (2019). Elucidating the expression and function of Numbl during cell adhesion-mediated drug resistance (CAM-DR) in multiple myeloma (MM). BMC Cancer, 19, 1269.

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