F12 medium

Manufactured by American Type Culture Collection

Sourced in United States

F12 Medium is a cell culture medium designed for the in vitro cultivation of a variety of cell types. It provides essential nutrients and supplements required for cell growth and maintenance.

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Close spelling variants of this product

DMEM/F12 (1:1) medium DMEM:F12 medium Dulbecco’s Modified Eagle’s Medium (DMEM-F12),

7 protocols using f12 medium

Alfalfa Flavonoids Impact on Cancer Cells

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Embryonic stem cell line (ESC) and endometrial carcinoma cell line (KLE) were all provided by the American Type Culture Collection (ATCC). ESC cells were cultured in 1:1 Mixture of Dulbecco’s Modified Eagles Medium and Ham’s F-12 medium (ATCC) supplemented with 2.0 mM L-Alanyl-L-Glutamine, 0.1 mM non-essential amino acids, 0.1 mM 2-mercaptoethanol, and 4 ng/ml bFGF as well as 15% fetal bovine serum (FBS). KLE cells were cultured in F12 Medium (ATCC) which was added with 10% FBS. All cells were incubated in 95% air and 5% CO₂ at 37°C.
AF (purity >98.0%) and N-acetyl cysteine (NAC, ROS inhibitor) were all purchased from Sigma-Aldrich (USA). ESC cells and KLE cells were treated with AF at different concentrations (50, 75, and 100 μM) for 48 h [15 (link)].
In another experiment, KLE cells were pre-treated with NAC (2.5 mM) for 30 min [16 (link)] and then treated with 100 μM AF for 48 h.

Sun Q., Zhen P., Li D., Liu X., Ding X, & Liu H. (2022). Amentoflavone promotes ferroptosis by regulating reactive oxygen species (ROS) /5’AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) to inhibit the malignant progression of endometrial carcinoma cells. Bioengineered, 13(5), 13269-13279.

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Osteoblastic Cell Culture with Graphene Composites

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Human fetal osteoblastic cells (hFOB 1.19) were cultured in a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and F-12 Medium (Kaighn's Modification of Ham's F-12 Medium; ATCC, Manassas, VA, USA). The culture medium was supplemented with 10% Fetal Bovine serum (FBS), 0.3mg/ml G418 (geneticin) and 1% Penicillin/Streptomycin (ATCC, Manassas, VA, USA). The hFOB 1.19 cells were plated in a six-well plate at a concentration of 1.0×10⁵ cells/well. The cells were allowed to reach 50-80% confluency after which the Gr-HA and CNTs-HA were added in duplicate to the cell culture medium in 200 and 400 ng/ml dosages. Control cells were grown in cell culture medium alone. The plates were incubated at 34°C and 5% CO₂. The day of plating was considered as day 0 of the experiment and the treatment medium renewed every 2-3 days.

Oyefusi A., Olanipekun O., Neelgund G.M., Peterson D., Stone J.M., Williams E., Carson L., Regisford G, & Oki A.A. (2014). Hydroxyapatite grafted carbon nanotubes and graphene nanosheets: promising bone implant materials. Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy, 132, 410-416.

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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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SH-SY5Y Cell Culture Protocol

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SH-SY5Y cells (cat. no. CRL2266, American Type Culture Collection, Manassas, VA, USA) were cultured in a 1:1 mixture of Eagle’s minimum essential medium (cat. no. 302003, ATCC) and F12 medium (cat. no. 302004, ATCC) supplemented with 10% fetal bovine serum (cat. no. SH30088.03, GE Healthcare Life Sciences, Pittsburgh, PA, USA).

Urasaki Y., Beaumont C., Workman M., Talbot J.N., Hill D.K, & Le T.T. (2020). Fast-Acting and Receptor-Mediated Regulation of Neuronal Signaling Pathways by Copaiba Essential Oil. International Journal of Molecular Sciences, 21(7), 2259.

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Cell Culture Protocols for Cancer Research

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Human colon epithelial cell line FHC (ATCC; Manassas, VA, USA) was routinely grown in DMEM: F12 Medium (ATCC). Human umbilical vein endothelial cells (HUVECs; ATCC) were maintained in F-12K Medium (ATCC). Besides, the colorectal cancer cell line SW480 (ATCC) was cultured in ATCC-formulated Leibovitz’s L-15 Medium. LoVo (ATCC) was cultured in ATCC-formulated F-12 K Medium. RKO (ATCC) was cultured in ATCC-formulated Eagle’s Minimum Essential Medium. HCT116 (ATCC) was cultured in ATCC-formulated McCoy’s 5a Medium Modified. Fetal bovine serum (FBS, 10%) was procured from Gibco (Rockville, MD, USA) and used for cell culture in a humidified atmosphere containing 5% CO₂ at 37 °C. Cells were passaged when the confluence was more than 80%.

Liu Y., Li Q., Tang D., Li M., Zhao P., Yang W., Shu L., Wang J., He Z., Li Y, & Wang F. (2020). SNHG17 promotes the proliferation and migration of colorectal adenocarcinoma cells by modulating CXCL12-mediated angiogenesis. Cancer Cell International, 20, 566.

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Culturing PC-3 Prostate Cancer Cells

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The human PC-3 PC cell line was purchased from
the American Type Culture Collection (ATCC CRL-1435). PC-3 cells were
cultured in F-12 medium (ATCC 30-2006) supplemented with 10% fetal
bovine serum and were grown to 80–90% confluence in 75 cm² culture flasks (Corning) for about a week (5–6 days)
in a humidified incubator (Fisher Scientific Isotemp) with 5% CO₂ at 37 °C. Culturing and subculturing procedures were
followed according to the ATCC Protocol available online for the PC-3
PC cell line.⁶⁹

Isaac-Lam M.F, & Hammonds D.M. (2019). Synthesis and Photodynamic Activity of Vitamin–Chlorin Conjugates at Nanomolar Concentrations against Prostate Cancer Cells. ACS Omega, 4(26), 21712-21723.

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Time-Course Cell Viability Assay in Serum/Growth Factor Deprivation

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To perform the time-course cell viability assay in the setting of serum/growth factor deprivation, HUVECs were incubated in 100% HPC-CM or 100% vehicle serum-free medium for 24, 48, and 72 hours, respectively. For all subsequent experiments for assessing gene expression, cell viability, cell proliferation, and apoptosis, HUVECs were treated with 50% HPC-CM, i.e. 1:1 mix of HPC-CM and HUVECs culture medium consisting of F-12 medium (30–2004, ATCC, VA, USA), 10% FBS (S009B, Millipore, MA, USA), 0.1 mg/mL heparin (H3393, Sigma, MO, USA), and 0.05 mg/mL endothelial cell growth supplement (E2759, Sigma, MO, USA) for 48 hours. HUVECs were also treated with 1:1 mix of vehicle serum-free medium and HUVECs culture medium as a control. To inhibit colony stimulating factor 1 (CSF1) signaling, HUVECs were treated with 1:1 mix of HPC-CM and HUVECs culture medium in the presence or absence of 30nM pexidartinib (B5854, ApexBio Technology, TX, USA).

Lee S., Karns R, & Shin S. (2022). Mechanism of paracrine communications between hepatic progenitor cells and endothelial cells. Cellular signalling, 100, 110458.

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