Cos 1 cells

Manufactured by American Type Culture Collection

Sourced in United States, United Kingdom

COS-1 cells are a cell line derived from the kidney of the African green monkey (Cercopithecus aethiops). They are commonly used as a host cell line for the production and study of recombinant proteins, as well as in various cell-based assays and research applications.

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COS-1 (49 citations) COS-1 cells from African green monkey kidney (2 citations) COS-1 and HEK293T cells (2 citations)

38 protocols using cos 1 cells

Cell Culture Conditions for Cancer Research

Cited 2 times

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EO771 and NCI-H1373 cells (ATCC) were maintained in RPMI (1.0 g/L glucose), supplemented with 10% v/v heat-inactivated fetal bovine serum (FBS), 1% v/v L-glutamine, and 1% v/v penicillin–streptomycin (P/S). COS-1 cells (ATCC) and MEFs (described elsewhere [16 (link),24 (link)]) were maintained in DMEM (1.0 g/L glucose) supplemented with 10% v/v FBS, 1% v/v L-glutamine, and 1% v/v P/S. Cells were cultured at 37 °C with 100% humidity and 5% CO₂. When confluency reached 80%, cells were subcultured. Cells were seeded at a density of 1 × 10⁵ cells per mL cell media 24 h prior to experiments unless otherwise stated.

Rasmussen M., Alvik K., Kannen V., Olafsen N.E., Erlingsson L.A., Grimaldi G., Takaoka A., Grant D.M, & Matthews J. (2023). Loss of PARP7 Increases Type I Interferon Signaling in EO771 Breast Cancer Cells and Prevents Mammary Tumor Growth by Increasing Antitumor Immunity. Cancers, 15(14), 3689.

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Isolation and Characterization of H9N2 Virus

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The H9N2 virus A/Chicken/Jiangsu/X1/2004 (X1) was isolated in Jiangsu Province, China. The virus was propagated in 9-day-old embryonated chicken eggs and preserved at − 70 °C. 1G8 hybridoma cells were screened from sp2/0 cells fused with spleen cells from BALB/C mice immunized with X1 virus, as previously reported (Wan et al. 2016 (link)). Madin-Darby canine kidney (MDCK) cells (ATCC, CCL-34) and COS-1 cells (ATCC, CRL-1650) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Thermo Fisher Scientific, Massachusetts, USA) supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Massachusetts, USA).

Wang F., Wang Y., Wan Z., Shao H., Qian K., Ye J, & Qin A. (2020). Generation of a recombinant chickenized monoclonal antibody against the neuraminidase of H9N2 avian influenza virus. AMB Express, 10, 151.

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Culturing of Diverse Cancer Cell Lines

Cited 1 time

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Human ZR75-1, MCF7, BT-474, T47D, MDA-MB-453, Hs578T, BT-549, MDA-MB-231 (American Type Culture Collection [ATCC, Manassas, VA]), and SUM159 breast carcinoma cells (from O. Kallioniemi, Institute for Molecular Medicine Finland, Helsinki, Finland; Neve et al., 2006 (link)), DU145 and PC3 prostate adenocarcinoma cells (ATCC), and WM852 melanoma cells (established at the Wistar Institute, Philadelphia, PA; Airola et al., 1999 (link)), and COS-1 cells (ATCC) were cultured according to manufacturer's instructions and as described (Tatti et al., 2011 (link)). Stable WM852 cell pools were generated by G418 (400 μg/ml; Calbiochem, Merck KGaA, Darmstadt, Germany) selection.

von Nandelstadh P., Gucciardo E., Lohi J., Li R., Sugiyama N., Carpen O, & Lehti K. (2014). Actin-associated protein palladin promotes tumor cell invasion by linking extracellular matrix degradation to cell cytoskeleton. Molecular Biology of the Cell, 25(17), 2556-2570.

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COS-1 Cell Culture in DMEM with FBS

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COS-1 cells (ATCC) were grown in DMEM supplemented with 10% (vol/vol) fetal bovine serum (FBS) (Invitrogen).

Sun J., Zhu G., Liu Y., Standley S., Ji A., Tunuguntla R., Wang Y., Claus C., Luo L., Baudry M, & Bi X. (2015). UBE3A regulates synaptic plasticity and learning and memory by controlling SK2 channel endocytosis. Cell reports, 12(3), 449-461.

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Transient Transfection of α1-Adrenergic Receptors

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COS-1 cells (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing glutamine and 5% fetal bovine serum. Cells were transiently transfected with purified plasmid DNA encoding WT or mutant α₁-ARs using FuGENE HD (Promega) following the manufacturer’s protocol. 48 h post-transfection, cells were harvested and homogenized using a Polytron homogenizer (Brinkmann Instruments) in HEM buffer (20 mM HEPES, 1.5 mM EGTA, 12.5 mM MgCl₂, pH 7.4) containing complete protease inhibitor (Roche Diagnostics). The homogenate was centrifuged at 484 × g (2000 rpm) for 10 min and the resulting supernatant was centrifuged at 23,665 × g (14000 rpm) for 30 min. The pellet was resuspended in HEM buffer containing 10% v/v glycerol and stored at −80°C prior to use. Protein concentration was determined using the BCA protein assay kit (Pierce) following the manufacturer’s protocol.

Dutt M., Giacomotto J., Ragnarsson L., Andersson Å., Brust A., Dekan Z., Alewood P.F, & Lewis R.J. (2019). The α1-adrenoceptor inhibitor ρ-TIA facilitates net hunting in piscivorous Conus tulipa. Scientific Reports, 9, 17841.

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Cell Culture and Transfection Protocol

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All cell lines were grown in a humidified incubator at 37°C and 5% CO2. COS-1 cells (ATCC CRL-1650) were maintained in Dulbecco’s modified Eagle’s medium (Gibco cat # 11995-065) supplemented with 10% bovine calf serum (HyClone cat# SH30072.03), 1% penicillin/streptomycin, and 1× GlutaMAX (Thermo Fisher Scientific). HEK293T cells were maintained similarly but in medium containing Fetal Clone III serum product (Hyclone cat # SH30109.03) instead of bovine calf serum (DuBridge et al. 1987 (link)). Cells were split and seeded onto poly-l-lysine coated coverslips in 35 mm petri dishes at 10% confluence and then transfected in Opti-MEM (Thermo Fisher Scientific) 24 hours later with 1 µg of the appropriate plasmid construct using LipofectAMINE 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol. Media was replaced 4 hours after transfection. Cells were grown for 40–48 hours post-transfection.

Speca D.J., Trimmer J.S., Peterson A.S, & Díaz E. (2017). Whole exome sequencing reveals a functional mutation in the GAIN domain of the Bai2 receptor underlying a forward mutagenesis hyperactivity QTL. Mammalian genome : official journal of the International Mammalian Genome Society, 28(11-12), 465-475.

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Transient and Stable Transfection of KCNC3 Variants

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COS-1 cells (ATCC, CRL 1650) were cultured in Dulbecco's Modified Eagle's Medium (DMEM, Omega) with 10% FBS (Omega), 4 mM glutamine, and antibiotic/antimycotic (ABAM) (Gibco) at 37°C in humidified air with 5% CO₂. Human neuroblastoma cells, SH-SY5Y (ATCC, CRL 2266) were cultured similarly in 1:1 Eagle's Minimum Essential Medium: F12 Medium (ATCC, 30-2003). The human KCNC3^WT cDNA was kindly provided by Dr. James L. Rae (Mayo Foundation) (Rae and Shepard, 2000 (link)) and subcloned into pcDNA1 (Invitrogen). KCNC3^WT and individual mutants (KCNC3^R420H or KCNC3^F448L) were generated by Quikchange Mutagenesis (Stratagene) and used for transient transfection in COS-1 cells. For co-transfection, equimolar amounts (1:1) of each plasmid were used. cDNAs for KCNC3^WT, KCNC3^R420H or KCNC3^F448L were subcloned into a pEF-IRES-puro vector for stable selection in SH-SY5Y cells using puromycin and individual colonies selected for clonal expression. Transfections were performed using Lipofectamine LTX (Invitrogen) as per manufacturer's instructions.

Gallego-Iradi C., Bickford J.S., Khare S., Hall A., Nick J.A., Salmasinia D., Wawrowsky K., Bannykh S., Huynh D.P., Rincon-Limas D.E., Pulst S.M., Nick H.S., Fernandez-Funez P, & Waters M.F. (2014). KCNC3R420H, a K+ Channel Mutation Causative in Spinocerebellar Ataxia 13 Displays Aberrant Intracellular Trafficking. Neurobiology of disease, 71, 270-279.

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Cos-1 Cell Transient Transfection

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Cos-1 cells (ATCC) were grown in DMEM supplemented with 10% (v/v) fetal bovine serum (FBS) (Invitrogen). For transient expression, cells were transfected with the respective DNA constructs by Lipofectamine 2000 (Invitrogen).

Wang Y., Hall R.A., Lee M., Kamgar-parsi A., Bi X, & Baudry M. (2017). The tyrosine phosphatase PTPN13/FAP-1 links calpain-2, TBI and tau tyrosine phosphorylation. Scientific Reports, 7, 11771.

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Cell Culture Transfection Experiments

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COS-1 cells (ATCC CRL-1650TM) were grown on DMEM (high glucose) without antibiotics supplemented with 10% fetal bovine serum at 37 °C under 5% CO₂. HeLa cells (ATCC CCL-2TM) were grown at 37 °C under 5% CO₂ on RPMI without antibiotics supplemented with 5% heat inactivated fetal bovine serum. All the transfection experiments were performed using Polyfect Reagent (Qiagen) following manufacturer’s instructions. For western blot experiments, cells were plated and transfected 24 h later with the appropriated plasmid. For luciferase experiments 3 × 10⁵ HeLa cells/well were plated on six wells dishes and transfected 24 h later with the appropriate concentrations of the specified plasmid. All the experiments were performed in triplicate and replicated twice. The quantity of plasmid for each well was kept constant by adding appropriate amounts of the corresponding empty plasmid.

Spinelli G., Biddeci G., Artale A., Valentino F., Tarantino G., Gallo G., Gianguzza F., Conaldi P.G., Corrao S., Gervasi F., Aronica T.S., Di Leonardo A., Duro G, & Di Blasi F. (2021). A new p65 isoform that bind the glucocorticoid hormone and is expressed in inflammation liver diseases and COVID-19. Scientific Reports, 11, 22913.

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COS-1 Cell Culture Protocol

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COS-1 cells (ATCC) were grown in DMEM supplemented with 10% (vol/vol) fetal bovine serum (FBS) (Invitrogen) and kept at 37°C under 5% CO₂. Low-passage cells were used, and the identity of cells has been authenticated by morphology check by microscope and growth curve analysis. No mycoplasma contamination was detected.

Sun J., Liu Y., Jia Y., Hao X., Lin W.J., Tran J., Lynch G., Baudry M, & Bi X. (2018). UBE3A-mediated p18/LAMTOR1 ubiquitination and degradation regulate mTORC1 activity and synaptic plasticity. eLife, 7, e37993.

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