Mouse BV-2 microglial cells (Peking Union Medical College Cell Bank, Beijing, China) were cultured in Dulbecco’s Modified Eagle’s Medium (Macgene, Beijing, China) and supplemented with 10% fetal bovine serum (Gibico, Waltham, MA, USA), penicillin (Macgene, 100 U/mL, Beijing, China), and streptomycin (Macgene, 100 μg/mL) in a humidified incubator containing 95% air and 5% CO2 at 37 °C.
Dulbecco s modified eagle s medium
Manufactured by Macgene
Sourced in China
Dulbecco's modified Eagle's medium (DMEM) is a cell culture medium that provides essential nutrients, growth factors, and other components to support the growth and maintenance of various cell lines in vitro. It is a widely used basal medium that can be supplemented with additional reagents as required for specific cell culture applications.
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Lab products found in correlation
Penicillin, by Macgene (2 mentions)
Streptomycin, by Macgene (2 mentions)
Rpmi 1640 medium, by Macgene (2 mentions)
Fetal bovine serum (fbs), by Macgene (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
B16 f10 luc, by American Type Culture Collection (1 mentions)
Mouse anti myc, by Santa Cruz Biotechnology (1 mentions)
Mccoy s 5a medium, by Macgene (1 mentions)
Rabbit anti dvl2, by Cell Signaling Technology (1 mentions)
Ic261, by Selleck Chemicals (1 mentions)
D4476, by Bio-Techne (1 mentions)
Mouse anti γ tubulin, by Merck Group (1 mentions)
Mouse anti acetylated tubulin, by Merck Group (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by R&D Systems (1 mentions)
Epidermal growth factor (egf), by R&D Systems (1 mentions)
6 protocols using dulbecco s modified eagle s medium
1
Culturing Mouse BV-2 Microglial Cells
2
Cell Culture Conditions for Cancer and Immune Cell Lines
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Human embryonic kidney 293T (HEK 293T) cells were purchased from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences (Beijing, China). Mouse dendritic (DC 2.4) cells were purchased from BeNa Culture Collection (Xinyang, Henan, China). Mouse melanoma cell lines (B16F10, B16F10-Luc, B16F10-OVA) and breast cancer 4T1 cells were purchased from American Type Culture Collection (ATCC, USA). HEK 293T cells, melanoma cells and DC 2.4 cells were cultured in Dulbecco's modified Eagle's medium (Macgene, Beijing, China) at 37 °C under 5 % CO2. 4T1 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Macgene, Beijing, China) at 37 °C under 5 % CO2. All these culture media were supplemented with 10 % fetal bovine serum (FBS; PAN, Beijing local agent, Germany), 100 U/mL penicillin, and 100 μg/mL streptomycin (Macgene, Beijing, China).
3
Cellular Signaling Pathway Investigation
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HEK293T, MCF7 and HeLa cells were cultured in Dulbecco’s modified Eagle’s medium (Macgene). HCT116 cells were cultured in McCoy’s 5A medium (Macgene). hTERT-RPE1 cells were cultured in DMEM/F12 medium (Gibco). Penicillin, streptomycin and 10% foetal bovine serum were added to all culture media unless otherwise noted. D4476 was purchased from Tocris Bioscience. IC261 was purchased from Selleckchem. Rabbit anti-Dvl2 and rabbit anti-PP2A/C antibodies were purchased from Cell Signaling Technology. Mouse anti-Dvl2 (10B5) and mouse anti-Myc antibodies were purchased from Santa Cruz Biotechnology. Rabbit anti-Dvl2 (phospho S143) and rabbit anti-γ-tubulin antibodies were purchased from Abcam. Mouse anti-PP5 antibody (for Western blotting) was purchased from BD Transduction Laboratories. Rabbit anti-PP5 antibody (for immunofluorescence) was purchased from Bethyl. Mouse anti-FLAG, mouse anti-acetylated tubulin and mouse anti-γ-tubulin antibodies were purchased from Sigma-Aldrich. Mouse anti-β-actin antibody was purchased from Sungene.
4
Maintenance of Cell Lines in Vitro
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Cell line PLC/PRF/5 and its derivatives were routinely maintained in Dulbecco’s modified Eagle’s medium (MACGENE Technology Ltd., Beijing, China) supplemented with 10% fetal bovine serum (Kangyuan Biology, China), and 100 units/mL penicillin plus 100 µg/mL streptomycin (Invitrogen). NK92MI cell line was maintained in RPMI 1640 medium (MACGENE Technology Ltd., Beijing, China) supplemented with 12.5% fetal bovine serum and 12.5% horse serum (Kangyuan Biology, China). Cell line CCRF-CEM and its derivative lines were maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum, and 100 units/mL penicillin plus 100 µg/mL streptomycin. All cells were cultured in the humidified incubator of 5% CO2 at 37 °C.
5
Cell culture of C6 and NIH 3T3 cell lines
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The rat glioma cell line (C6) and the mouse embryonic fibroblasts cell line (NIH 3T3) were obtained from the Institute of Basic Medical Sciences (Chinese Academy of Medical Sciences, Beijing, People’s Republic of China) and cultivated under the recommended conditions. Briefly, the C6 cells were maintained in Ham’s F10 Medium (Macgene Biotech Co., Ltd., Beijing, People’s Republic of China) supplemented with 15% (v/v) horse serum (Thermo Fisher Scientific, Waltham, MA, USA), 2.5% (v/v) fetal bovine serum (FBS, Thermo Fisher Scientific), 100 U/mL penicillin, and 100 μg/mL streptomycin. The NIH 3T3 cells were cultured in Dulbecco’s Modified Eagle’s Medium (Macgene Biotech Co., Ltd., Beijing, People’s Republic of China) supplemented with 10% (v/v) fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin. Cells were seeded into plates precoated with 0.02% (w/v) gelatin in a 5% CO2-humidified chamber at 37°C, with a medium change at the first 24 hours and every 48 hours thereafter until the cells reached approximately 80%–90% confluence. The cells were then trypsinized and transferred into a gelatin-coated culture flask with culture medium for the follow-up study.
6
Glioblastoma Stem Cell Differentiation
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GSCs (specimens 387 and 4121) were originally isolated and characterized from human GBM surgical specimens as previously described (26 (link)) and cultured as neurospheres in neurobasal complete media (Gibco) supplemented with 1× B27 (Gibco) (without vitamin A), 2 mM l-glutamine (MacGene), 1 mM sodium pyruvate (MacGene), 10 ng/ml basic fibroblast growth factor (R&D Systems), and 10 ng/ml epidermal growth factor (R&D Systems). DGCs were differentiated from GSCs via the withdrawal of epidermal growth factor and fibroblast growth factor and addition of 10% fetal bovine serum (FBS; MacGene). DGCs were cultured in Dulbecco’s modified Eagle’s medium (MacGene) containing 10% FBS. Expression of the GSC marker SOX2/Olig2 and the differentiation marker GFAP was characterized using Western blotting or immunofluorescence staining. All cells were incubated at 37°C in a humidified incubator supplemented with 5% CO2.
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