Mccoy s 5a medium

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About the product

McCoy's 5A medium is a cell culture medium used for the growth and maintenance of various cell types. It provides the necessary nutrients and growth factors required for cell proliferation and survival. The medium's composition is designed to support the specific needs of the targeted cell lines.

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McCoy's 5A (Modified) Medium is a cell culture medium commercially available from Thermo Fisher Scientific. The product is currently in production and can be purchased through authorized distributors.

Prices for McCoy's 5A (Modified) Medium typically range from $35.65 to $61.50 per 500 mL bottle, depending on the specific formulation and supplier.

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McCoy’s 5A (595 citations) McCoy’s medium (159 citations) McCoy 5A medium (76 citations) McCoy 5A (52 citations) McCoy medium (37 citations) McCoy’s (30 citations) ’s medium (1 citations)

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1 755 protocols using mccoy s 5a medium

Colorectal Cancer Cell Line Cultivation

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CRC cell lines of SW480, HCT116, SW620, and DLD1 were supplied by Cell Bank, Chinese Academy of Sciences. Human normal colonic epithelial cells NCM460 were purchased from the American Type Culture Collection (Manassas, Virginia, USA). SW480 and SW620 cells were cultured in L‐15 (11415064, Gibco) medium containing 1% penicillin/streptomycin and 10% fetal bovine serum (Gibco, USA) at 37°C in an incubator. HCT116 cells were cultured in McCoy's 5a medium (12330031, Gibco) supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum at 37°C in a 5% CO₂ incubator. DLD1 cells were cultured in RPMI 1640 medium (11875093, Gibco) supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum at 37°C in a 5% CO₂ incubator. NCM460 cells were cultured in Dulbecco's modified Eagle's medium (KeyGEN BioTECH, Jiangsu, China) with 10% fetal bovine serum (Gibco, USA) at 37°C in a 5% CO₂ incubator.
For experimental purposes, cells were transfected with overexpression vectors to upregulate target gene expression, siRNA to knockdown specific gene expression, block oligonucleotides to inhibit mRNA translation, and a negative control to ensure the specificity of the observed effects. All transfections were performed using the Lipo6000 transfection reagent, following the manufacturer's protocols for transient transfection.

Ma Z., Zhu J., Chen M., Wu G., Liu X., Hu Z., Feng Y., Wang X, & Liu F. (2025). RALYL Overexpression Suppresses Colorectal Cancer via Modulating HNRNPC‐Mediated MNK2 Alternative Splicing. Cancer Reports, 8(3), e70179.

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Establishing and Characterizing Human Cervical and Head-Neck Cancer Cell Lines

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Human cervical cancer cell lines C-33A, C-4 I, CaSki, DoTc2 4510, HeLa, MS751, and SiHa were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) formulated with high glucose and l-glutamine (Thermo Fisher Scientific, Waltham, MA) and supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (all from Thermo Fisher Scientific). Human cervical cancer cell line HT-3 was obtained from the ATCC and cultured in McCoy’s 5a medium supplemented with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin (all from Thermo Fisher Scientific). Two cervical cancer cell lines, 778 and 879, were previously provided by Dr. P.L. Stern (Paterson Institute for Cancer Research, Manchester, UK) and cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% FBS, penicillin at 100 U/mL, and streptomycin at 100 μg/mL (all from Thermo Fisher Scientific).⁷³ (link) Head and neck squamous cell carcinoma (HNSCC) cell lines UM-SCC-47, VU-SCC-040, VU-SCC-120, and VU-SCC-147 were obtained as described previously and cultured with DMEM supplemented with 10% FBS, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 μg/mL streptomycin (all from Thermo Fisher Scientific).⁷⁴ (link)^,⁷⁵ (link)^,⁷⁶ (link) Cell cultures were maintained at 37°C in a humidified atmosphere with 5% CO₂. Cell lines were authenticated by short-tandem repeat (STR) testing, monthly tested for the presence of mycoplasma and HPV status was confirmed using the QIAscreen HPV PCR assay (Qiagen, Hilden, Germany).

Xu M., van de Wiel M.A., Martinovičová D., Huseinovic A., van Beusechem V.W., Stalpers L.J., Oei A.L., Steenbergen R.D, & Snoek B.C. (2025). High-throughput 3D spheroid screens identify microRNA sensitizers for improved thermoradiotherapy in locally advanced cancers. Molecular Therapy. Nucleic Acids, 36(2), 102500.

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Bladder Cell Line Culture and Treatments

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The bladder cell lines T24 cells and 5637 cells were purchased from Stem Cell Bank (Chinese Academy of Sciences, Beijing, China) and cultured in McCoy’s 5 A medium (Thermo Fisher Scientific, Waltham, MA, USA, catalog no. 16600082) or RPMI 1640 medium (Thermo Fisher Scientific, Waltham, USA catalog no. 11875093) with 10% fetal bovine serum (FBS, AusGeneX, Gold Coast, Australia, catalog no. FBS500-S). Oxaliplatin (OXA, catalog no. S1224), SRT3025 (catalog no. S8481), and compound 3 K, the inhibitor of PKM2, (3 K, catalog no. S8616), were purchased from Selleck Chemicals (Houston, USA).

Wang X., Li S., Li Z., Lin Z, & Wang Z. (2025). SRT3025-loaded cell membrane hybrid liposomes (3025@ML) enhanced anti-tumor activity of Oxaliplatin via inhibiting pyruvate kinase M2 and fatty acid synthase. Lipids in Health and Disease, 24, 14.

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Screening Compound Library for p53 Activation

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Cell culture. HCT116, human colorectal carcinoma cell line, were cultured in McCoy’s 5A medium (Gibco) supplemented with 10% fetal bovine serum (FBS). HCT116 was obtained from the Cell Bank of the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). The cells were maintained in an incubator at 37 °C with 5% CO\documentclass[12pt]{minimal}
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Luciferase activity assay. A luciferase activity assay was performed as described previously³⁰. Briefly, cells containing the p53-driven luciferase reporter gene were plated in 96-well plates at a density of \documentclass[12pt]{minimal}
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\begin{document}$$1 \times 10^{4}$$\end{document} cells/well. Twelve hours after the initial incubation, the test compounds (a library containing 4000 compounds) were added to each well at specific concentrations, while an equal volume of the solvent DMSO was used as a blank control. Luciferase activity was measured via the Luciferase Assay System (Cat#E2920, Promega) on a Molecular Devices/SpectraMaxL instrument with a 562 nm emission filter set.

Wang X., Zhang M., Xu J., Li X., Xiong J., Cao H., Dou F., Zhai X, & Sun H. (2025). A novel approach for target deconvolution from phenotype-based screening using knowledge graph. Scientific Reports, 15, 2414.

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Ovarian Cancer Cell Lines Knockdown

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Human ovarian epithelial cells IOSE-80 (BNCC358126) as well as OC cell lines SK-OV-3 (BNCC338639), CoC1 (BNCC101678), A2780 (BNCC351906) and HEY (BNCC353326) were all ordered from BeiNa Culture Collection (Xinyang, China) High-glucose Dulbecco’s modified Eagle’s medium (11,965,092, Gibco, Grand Island, NY), Roswell Park Memorial Institute-1640 medium (11875101, Gibco) and McCoy’s 5A medium (16600082, Gibco) were used in cell culture. 10% fetal bovine serum (A3161001C, Gibco) was supplemented in all the media. Cell incubation was performed using an incubator at 37 ℃ with 5% CO₂ in a humidified atmosphere. Lipofectamine 2000 (11668027) was procured from Invitrogen (Carlsbad, CA) and the relevant small interfering RNAs (siRNAs) targeting FOXJ2 si-FOXJ2#1 (target sequence: 5′-GTGATAACTTCCCCTATTACAAG-3′) and si-FOXJ2#2 (target sequence: 5′-GGGTTCCTATTGGACAATTGACA-3′) as well as the control siRNA (si-NC) with the scrambled sequence were all ordered from RiboBio (Guangzhou, China). Then OC cells A2780 and HEY were transfected with the relevant siRNAs with the transfection reagent according to the protocol of the producer. All cells have been STR identified and the mycoplasma detection results are negative.

Wang L., He H., Zhai R., Gao R., Su M., Duan R., Tu Z, & Huang R. (2025). Investigation of the mechanism by which FOXJ2 inhibits proliferation, metastasis and cell cycle progression of ovarian cancer cells through the PI3K/AKT signaling pathway. European Journal of Medical Research, 30, 152.

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Human Cancer Cell Line Cultivation

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The human cancer cell lines utilized in this study, obtained from the Cell Bank of the Chinese Academy of Sciences and maintained in our laboratory, were all STR profiled upon entry into the repository. SK-OV-3 and HEC-1-A cells were cultured in McCoy’s 5A medium (Gibco, USA), A2780 cells in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, USA), and A549, PANC-1, Ishikawa, HeLa, and SiHa cells in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, USA). Each medium was supplemented with 10% fetal bovine serum (FBS, Vazyme, China) and 1% penicillin-streptomycin (Servicebio, China). Cells were incubated at 37 °C in a humidified incubator containing 5% CO2. The culture medium was refreshed every 3–4 days, and cells were subcultured once they reached 80–90% confluence. All procedures were conducted under sterile conditions, and regular mycoplasma testing was performed to ensure the validity of the experimental data.

Li W., Ji T., Ye J., Xiong S., Si Y., Sun X., Li F, & Dai Z. (2025). Ferroptosis enhances the therapeutic potential of oncolytic adenoviruses KD01 against cancer. Cancer Gene Therapy, 32(4), 403-417.

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Establishing Fluorescent Cell Lines

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The human renal clear cell adenocarcinoma 786-0, glioma U251, pancreatic adenocarcinoma Capan-2, triple-negative breast cancer HCC1806 cells were obtained from the American Type Culture Collection (ATCC). Stable cell lines expressing green fluorescent protein (GFP) and firefly luciferase (ffLuc) were generated through lentiviral transduction. CD70 knockout 786-0 cells (CD70 KO 786-0) were generated using the CRISPR/Cas9 system. The 786-0 and 1806 cells were cultured in RPMI 1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco). U251 cells were cultured in DMEM medium (Gibco) with 10% FBS. Capan-2 cells were cultured in McCoy’s 5a medium (Gibco). All cells were maintained in a 37°C incubator with 5% CO₂.

Yin X., Chen W., Ao X., Xu L., Cao J., Huang T., Liang J., Hu J., Liu J., Wang X., Li W., Zhou M., He L, & Guo Z. (2025). Sodium citrate pretreatment enhances CAR-T cell persistence and anti-tumor efficacy through inhibition of calcium signaling. Frontiers in Immunology, 16, 1540754.

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Cell Culture Protocols for Cancer Research

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RKO, HCT116, MC38, HIEC-6, and 293T cell lines were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All cell lines were identified by Short Tandem Repeat profiling by the source. RKO, MC38, and 293T cells were cultured in DMEM medium (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin-streptomycin (100 U/mL penicillin and 100 μg/mL streptomycin). HCT116 cells were cultured in McCoy’s 5A medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin-streptomycin (Gibco, USA). All cells were maintained at 37 °C in a humidified atmosphere containing 5% CO₂.

Gao C., Chen L., Zhao L., Su Y., Ma M., Zhang W., Hong X., Xiao L., Xu B, & Hu T. (2025). Apatinib Degrades PD-L1 and Reconstitutes Colon Cancer Microenvironment via the Regulation of Myoferlin. Cancers, 17(3), 524.

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Ovarian Cancer Cell Culture Conditions

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All cell lines were acquired from the American Type Culture Collection (Manassas, VA, USA) and maintained using recommended culture conditions. The OVCAR-3 (HTB-161) ovarian cancer cell line was maintained in RPMI 1640 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 20% (v/v) fetal bovine serum (FBS; Bovogen, Keilor East, VIC, Australia), 0.01 mg/mL bovine insulin (Sigma-Aldrich), and 1× penicillin-streptomycin (Pen/Strep; Gibco, Waltham, MA, USA). The ovarian cancer cell line derived from ascites, MES-OV (CRL-3272), was cultured in McCoy’s 5A medium (Gibco) containing 10% (v/v) FBS and 1× Pen/Strep.

Evtimov V.J., Nguyen N.Y., Hammett M.V., Pupovac A., Hudson P.J., Zhuang J., Lee J.Y., Kim S., Trounson A.O., Boyd R.L, & Shu R. (2025). CRISPR-Cas9 knockout of DGKα/ζ improves the anti-tumor activities of TAG-72 CAR-T cells in ovarian cancer. Molecular Therapy Oncology, 33(2), 200962.

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Isolation of Extracellular Vesicles from Cell Lines

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Six cell lines were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). Normal human bladder urothelial cells (SVHUC1) were cultured in Ham's F-12K medium supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco) and 1% (v/v) penicillin-streptomycin (PS, Gibco) in an incubator under 5% CO₂ at 37 °C. RT4 was cultured in McCoy's 5A medium (Gibco) supplemented with 10% (v/v) FBS (Gibco) and 1% (v/v) PS (Gibco) in an incubator under 5% CO₂ at 37 °C. The HK-2, 5637, T24, and EJ cells were cultured in RPMI-1640 medium (Gibco) supplemented with 10% (v/v) FBS (Gibco) and 1% PS (v/v) (Gibco) in an incubator under 5% CO₂ at 37 °C. An appropriate volume of serum-free medium was added until the cells grew to 70-80% density. After 48 hours, the supernatant was collected for EVs isolation.

Sun N., Zhang Z., Yang X., Li J., Li Q., Kang J., Wei Y., Yu X., Du R., Hong X., Liu G., Gao H, & Liu D. (2025). Unveiling urinary extracellular vesicle mRNA signature for early diagnosis and prognosis of bladder cancer. Theranostics, 15(4), 1272-1284.

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