Fetal bovine serum (fbs)

Manufactured by Avantor

Sourced in United States, Canada, Germany, France, Australia, Panama, China, Sweden, Spain

FBS is a serum supplement commonly used in cell culture applications. It provides essential nutrients and growth factors to support the growth and proliferation of a variety of cell types.

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Lab products found in correlation

527 protocols using fetal bovine serum (fbs)

Cell Culture Conditions for 293T, hTert-BJ1, and U-937

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293T (CRL-3216), hTert-BJ1 (BJ-5ta - CRL-4001) and U-937 (CRL-1593.2) were from ATCC. 293T were cultured in DMEM (Corning) supplemented with 10% FBS (VWR), 1X GlutaMax (Thermo Fisher) and 1X Penicillin/Streptomycin (Corning). hTert-BJ1 were cultured in a 4:1 mixture of DMEM (Corning):Medium 199 (Lonza) supplemented with 10% FBS (VWR), 1X GlutaMax (Thermo Fisher) and 1X Penicillin/Streptomycin (Corning). U-937 and THP-1 were cultured in RPMI (Thermo Fisher) supplemented with 10% FBS (VWR), 1X GlutaMax (Thermo Fisher) and 1X Penicillin/Streptomycin (Corning).

Gentili M., Liu B., Papanastasiou M., Dele-Oni D., Schwartz M.A., Carlson R.J., Al’Khafaji A.M., Krug K., Brown A., Doench J.G., Carr S.A, & Hacohen N. (2023). ESCRT-dependent STING degradation inhibits steady-state and cGAMP-induced signalling. Nature Communications, 14, 611.

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Cell Culture Protocols for Cancer Research

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A20 and B16F10 cells were purchased from ATCC. OCI-LY3 and SU-DHL-4 were generously shared by Dr. Han Tun, with genomic profiling performed November 2022 (GENOMIC). A20 cells were cultured in RPMI (Fisher-Scientific) supplemented with 10% FBS (VWR) and 1% Penn-Strep (Life Technologies). OCI-LY3 and SU-DHL-4 cells were cultured in RPMI (Fisher-Scientific) supplemented with 20% FBS (VWR) and 1% Penn-Strep (Life Technologies). B16F10 cells were cultured in DMEM (Fisher-Scientific) supplemented with 10% FBS (VWR) and 1% Penn-Strep (Life Technologies). All cell lines were maintained at 37°C in humidified conditions with 5% CO₂. At the beginning of the study, cells were expanded, stocks made, and thawed vials were maintained in culture for no more than 3 weeks.

Von Roemeling C.A., Doonan B.P., Klippel K., Schultz D., Hoang-Minh L., Trivedi V., Li C., Russell R.A., Kanumuri R.S., Sharma A., Tun H.W, & Mitchell D.A. (2023). Oral IRAK-4 Inhibitor CA-4948 Is Blood-Brain Barrier Penetrant and Has Single-Agent Activity against CNS Lymphoma and Melanoma Brain Metastases. Clinical Cancer Research, 29(9), 1751-1762.

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Cell Line Culture Protocols

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The female human embryonic kidney cell line HEK293T (Cat#CRL-3216; RRID: CVCL_0063) was obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 4.5 g/L glucose, L-glutamine, and sodium pyruvate (Corning) supplemented with 10% (v/v) fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). Immortalized mouse hippocampal cell line mHippoE-2 (Cat# CVCL_D377; RRID: CVCL_D377) was obtained from Cedarlane Laboratories and cultured in DMEM with 4.5 g/L glucose, L-glutamine, and sodium pyruvate (Corning) supplemented with 10% (v/v) fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). The sex of mHippoE-2 cells was not disclosed by the manufacturer and may be a mixed population of male and female. The female human neuroblastoma cell line SH-SY5Y (Cat#CRL-2266; RRID: CVCL_0019) was obtained from the ATCC and cultured in equal parts Minimum Essential Medium Eagle (EMEM) with Earle′s salts, L-glutamine and sodium bicarbonate (Sigma-Aldrich) and Ham’s F12 Nutrient Mix (ThermoFisher), supplemented with 10% (v/v) gamma-irradiated and heat-treated fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). Cell lines used in this study have not been authenticated. Cell lines used in this study were cultured in a humidified 37°C incubator with 5% CO₂.

Choi S.H., Flamand M.N., Liu B., Zhu H., Hu M., Wang M., Sewell J., Holley C.L., Al-Hashimi H.M, & Meyer K.D. (2022). RBM45 is an m6A-binding protein that affects neuronal differentiation and the splicing of a subset of mRNAs. Cell reports, 40(9), 111293.

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Cell Line Culture Protocols

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Detailed Cell Culture Protocols

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HepG2 and Huh7 cells were obtained from ATCC. MDA-MB231 cells were obtained from Dr. Hasan Korkaya who originally purchased from ATCC. All cell lines were maintained in DMEM medium supplemented with 10% FBS (VWR). The Epstein-Barr virus (EBV)–transformed B-lymphoblastoid cell lines (B-LCL) used for alloreactivity test were obtained from either Sigma or Fred Hutchinson Cancer Research Center, and maintained in RPMI 1640 medium supplemented with 15% FBS (VWR). Primary adult human hepatocytes were obtained from Lonza. Primary human lung and kidney epithelial cells were purchased from Novabiosis and Lifeline, respectively. Induced pluripotent stem cell-derived iCell® Neurons, Astrocytes, Cardiomyocytes and Endothelial Cells were all from FUJIFILM Cellular Dynamics, Inc. The culture medium, supplements and plate coating reagents for each cell type were purchased from vendors as designated by the cell suppliers. Primary hepatocytes and the four iCells were originally from HLA-A^*02:01⁺ donors, according to vendor-provided information. Primary lung and kidney epithelial cells were originally from HLA-A2⁺ donors, in-house PCR (25 (link)) and sequencing further confirmed they also carry HLA-A^*02:01 allele.

Luo X., Cui H., Cai L., Zhu W., Yang W.C., Patrick M., Zhu S., Huang J., Yao X., Yao Y., He Y, & Ji Y. (2020). Selection of a Clinical Lead TCR Targeting Alpha-Fetoprotein-Positive Liver Cancer Based on a Balance of Risk and Benefit. Frontiers in Immunology, 11, 623.

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Culturing HEK293, HeLa, and U937 Cells

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HEK293T cells (female), HEK293 cells (female) stably expressing the Fcγ receptor IIb (HEK293 FcγR cells), and HeLa FcγR cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, GIBCO) containing 10% fetal bovine serum (FBS, VWR) at 37°C and 5% CO₂. FcγR expressing cell lines were gifts from the lab of Dr. Craig Roy at Yale University. U937 cells (a gift from Dr. Michael Bassik at Stanford University) were cultured in RPMI-1640 (Corning) supplemented with 10% heat-inactivated FBS (VWR). U937 were differentiated into macrophage-like cells in 20 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 72 hours, then re-plated in media without PMA and allowed to rest for 48 hours before L.p infection.

Steinbach A.M., Bhadkamkar V.L., Jimenez-Morales D., Stevenson E., Jang G.M., Krogan N.J., Swaney D.L, & Mukherjee S. (2023). Cross-family small GTPase ubiquitination by the intracellular pathogen Legionella pneumophila. bioRxiv.

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Isolation of Lymphocytes from Primate Samples

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Peripheral blood mononuclear cells were isolated from pigtail macaque blood by Histopaque-1077 (Sigma-Aldrich) density gradient in Accupsin conical tubes (Sigma-Aldrich). Red blood cells were lysed using ACK lysis buffer (ThermoFisher) and then resuspended in RPMI-1640 media (Invitrogen) supplemented with 10% Fetal Bovine Serum (FBS) (VWR), 50 µg/ml Gentamicin sulfate (ThermoFisher), and 5 mg/ml of Penicillin–Streptomycin–Glutamine (Pen–Strep–Glut) (ThermoFisher). Intraepithelial and lamina propria lymphocytes of gut and vaginal mucosa biopsies were isolated by straining through a 70-micron filter (Corning) following a 1 hour (h) enzymatic digestion at 37 °C with 40 µg/ml Liberase TM (Sigma-Aldrich) and 80 µg/ml DNase (Sigma-Aldrich) in RPMI-1640 media supplemented with 5 mg/ml of Pen–Strep–Glut (ThermoFisher). Lymphocytes from whole lymph node biopsies were isolated by directly straining through a 70-micron filter in RPMI-1640 media supplemented with 10% FBS (VWR) and 5 ml of 1X Pen–Strep–Glut (ThermoFisher), and 50 μg/ml Gentamicin sulfate (ThermoFisher).

O’Connor M.A., Tisoncik-Go J., Lewis T.B., Miller C.J., Bratt D., Moats C.R., Edlefsen P.T., Smedley J., Klatt N.R., Gale M J.r, & Fuller D.H. (2018). Early cellular innate immune responses drive Zika viral persistence and tissue tropism in pigtail macaques. Nature Communications, 9, 3371.

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Cultivation of T. cruzi Epimastigotes and Amastigotes

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Epimastigotes of T. cruzi Y strain were cultured in LDNT/LIT medium (Kirchhoff et al., 1984 (link)) supplemented with 15% FBS. Aliquots of FBS (VWR, USDA certified) were freshly heat inactivated at 76°C for 40 min (to help improve epimastigote viability at low numbers) prior to addition to the media (this heat inactivated FBS will precipitate if frozen). To obtain amastigotes, 1 mL of maximum density epimastigotes were differentiated to metacyclic trypomastigotes via prolonged starvation in depleted LIT media (for at least 3 days). The resulting parasites were then incubated overnight with fresh FBS (with intact complement) to kill any remaining epimastigotes. The surviving metacyclic trypomastigotes were added to flasks containing human foreskin fibroblasts (HFF) grown in High Glucose Dulbecco's Modified Eagle's Medium (DMEM-HG) (Hyclone) supplemented with L-glutamine and 10% heat-inactivated (56°C) Cosmic Calf serum (CCS) (Hyclone). After 24–48 h media was replaced with medium containing only 1% (CCS) for further culturing.

Chasen N.M., Coppens I, & Etheridge R.D. (2020). Identification and Localization of the First Known Proteins of the Trypanosoma cruzi Cytostome Cytopharynx Endocytic Complex. Frontiers in Cellular and Infection Microbiology, 9, 445.

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Cell Culture and Macrophage Differentiation

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HEK293T cells (female), HEK293 cells (female) stably expressing the Fcγ receptor IIb (HEK293 FcγR cells), and HeLa FcγR cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, GIBCO) containing 10% FBS (VWR) at 37°C and 5% CO₂. FcγR-expressing cell lines were gifts from the lab of Dr. Craig Roy at Yale University. U937 cells (a gift from Dr. Michael Bassik at Stanford University) were cultured in RPMI-1640 (Corning) supplemented with 10% heat-inactivated FBS (VWR). U937 were differentiated into macrophage-like cells in 20 ng/ml phorbol 12-myristate 13-acetate (Sigma) for 72 h, then replated in media without PMA and allowed to rest for 48 h before L.p infection.

Steinbach A., Bhadkamkar V., Jimenez-Morales D., Stevenson E., Jang G.M., Krogan N.J., Swaney D.L, & Mukherjee S. (2024). Cross-family small GTPase ubiquitination by the intracellular pathogen Legionella pneumophila. Molecular Biology of the Cell, 35(3), ar27.

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Culturing and Maintaining Cell Lines

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The B16/F10 cell line used for murine models was a CRISPR scramble control from the original ATCC cell bank. The BSC40 and Jurkat cell lines were purchased from ATCC (Manassas, VA). LLC-A9F1 cells were a gift from Dr. Mark Rubinstein at the Medical University of South Carolina. Jurkat cells were maintained in RPMI (Corning, Corning, NY) supplemented with 10% FBS (VWR, Radnor, PA) and 1x penicillin/streptomycin/glutamine. For Jurkat activation experiments, cells were treated with 5ng/mL PMA and 500ng/mL Ionomycin for 2 hours in RPMI, washed with PBS, and plated in fresh RPMI. All other cell lines were maintained in DMEM (Corning, Corning, NY) supplemented with 10% FBS (VWR, Radnor, PA) and 1x penicillin/streptomycin/glutamine (Corning, Corning, NY). Cultures were checked quarterly for mycoplasma contamination by qRT-PCR.

Dryja P., Fisher C., Woster P.M, & Bartee E. (2021). Inhibition of polyamine biosynthesis using difluoromethylornithine acts as a potent immune modulator and displays therapeutic synergy with PD1-blockade. Journal of immunotherapy (Hagerstown, Md. : 1997), 44(8), 283-291.

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