Murine B16F10 melanoma cells purchased from ATCC (Manassas, VA, USA) were cultured in DMEM with 10 % (v/v) fetal bovine serum and 1 % (v/v) antibiotic–antimycotic. The cells were incubated in a humidified incubator under 5 % CO2 at 37 °C. Aqueous fractions are dissolved in DDW and EtOAc fractions are dissolved in DMSO. Controls of B16F10 cells were added DDW or DMSO at the same concentration as in the treated cells.
B16f10 murine melanoma cells
Manufactured by American Type Culture Collection
Sourced in United States
B16F10 murine melanoma cells are an established cell line derived from a C57BL/6 mouse melanoma. They are widely used in cancer research as a model for studying melanoma.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Cytiva (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Cytiva (3 mentions)
Penicillin, by Cytiva (3 mentions)
Streptomycin, by Cytiva (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Female c57bl 6 mice, by Charles River Laboratories (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Dota tri t butyl ester, by Macrocyclics (2 mentions)
Isolink kit, by Mallinckrodt (2 mentions)
Synergy ht multi mode microplate reader, by Agilent Technologies (2 mentions)
L glutamine, by Corning (2 mentions)
Hdbec, by PromoCell (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Mycoalert detection kit, by Lonza (1 mentions)
63 protocols using b16f10 murine melanoma cells
1
Murine B16F10 Melanoma Cell Culture
2
Primary Endothelial Cell Culture Protocol
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Primary Human Dermal Blood Endothelial Cells (HDBEC) (PromoCell) and Human Umbilical Vein Endothelial Cells (HUVEC) (3 H Biomedical) were cultured in Endothelial Cell Growth Medium 2 (EMV2) (PromoCell) in gelatin-coated plates. Murine B16-F10 melanoma cells (ATCC CRL-6475, American Type Culture Collection) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with GlutaMAX (Thermofisher Scientific) and 10% fetal bovine serum (FBS) (Sigma-Aldrich). HCmel12 murine melanoma cells46 ,47 were generously provided by Prof. T. Tüting (Laboratory of Experimental Dermatology, University of Bonn, Germany) and cultured in RPMI 1640 Medium (Thermofisher Scientific) with 10% FBS. Cell cultures were incubated at 37°C with 5% CO2 in a humidified cell incubator. No authentication of cell lines has been performed after purchase. All cell cultures were routinely tested negative for mycoplasma contamination using the MycoAlert Detection Kit (Lonza, Basel, Switzerland).
3
Murine B16F10 Melanoma Tumor Implantation
4
Pinostrobin's Effect on Melanoma Cell Viability
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Murine B16F10 melanoma cells (ATCC, Manassas, VA, USA) were maintained in DMEM supplemented with 10% heat-inactivated FBS at 37 °C in a humidified atmosphere containing 5% CO2. To analyze the effect of pinostrobin on cell viability, an MTT assay was performed. Briefly, B16F10 melanoma cells were seeded in 24-well plates at a density of 1 × 104 cells/mL for 16 h. The cells were then treated with the indicated concentrations of pinostrobin (0–200 μM) for 24, 48, and 72 h. After incubation, MTT was added to each well and the plates were incubated for 4 h at 37 °C. The precipitate was dissolved in dimethyl sulfoxide (DMSO), and the absorbance was measured at 540 nm using a microplate spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Cellular morphology was observed under a stereomicroscope (MACROTECH, Goyang, Gyeonggi-do, Republic of Korea).
5
Murine B16F10 Melanoma Tumor Implantation
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Cell culture: Murine B16F10 melanoma cells (CRL-6475) were purchased from ATCC. Cells were cultured at 37°C in a humidified atmosphere of 5% CO2, high glucose (4.5 g/L) DMEM supplemented with 10% fetal bovine serum and 1% glutamine and were used by passage 10 for implantation into syngeneic recipient mice. Tumor cell implantation: 100 μL of B16F10 melanoma cells at 1.0 × 107/mL in Dulbecco’s phosphate-buffered saline (PBS) were injected subcutaneously into the back of 6- to 8-week-old Pf4-Cre Clec1bfl/fl female mice.
6
Culturing Murine Melanoma Cells
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Murine B16-F10 melanoma cells (ATCC, USA) were cultured in DMEM/10% (v/v) fetal calf serum/2 mM L-glutamine/100 U/ml penicillin/100 μg/ml streptomycin (all Life Technologies, Germany) at 37°C and 5% (v/v) CO2.
7
Cell Culture Conditions for Immunological Studies
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Human embryonic kidney-293 cells (ATCC) were cultured in Eagle’s Minimum Essential Medium containing 10% heat-inactivated bovine calf serum (GE Healthcare Life Sciences, USA). Murine B16-F10 melanoma cells (ATCC) that were used to establish tumors were cultured in Dulbecco’s Modified Eagle’s Medium (GE Healthcare Life Sciences) containing 10% bovine calf serum. Vero cells were used for an in-cell western blotting assay to quantify antibody responses. They were also grown in Dulbecco’s Modified Eagle’s Medium with 10% bovine calf serum. All cells were cultured at 37 °C in a humidified atmosphere with 5% CO2 and were confirmed to be mycoplasma-free prior to use (MycoAlert PLUS detection kit, Lonza, USA).
8
Murine B16F10 Melanoma Cell Assays
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Murine B16F10 melanoma cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and used for cell viability, cellular tyrosinase activity, and melanin content assays. B16F10 cells were cultured in Dulbecco’s modified Eagle’s medium containing 10% FBS (fetal bovine serum), and 1% streptomycin at 37 °C in a 5% CO2 environment, and these cells were then used in cell viability, cellular tyrosinase activity, and melanin content assays in 96-well plates or 6-well dishes. All experiments were independently performed in triplicate.
9
Culturing Murine Melanoma and Endothelial Cells
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Murine B16F10 melanoma cells and immortalized mouse brain endothelial cells, bEnd.3, were obtained from the American Type Culture Collection (Manassas, VA, United States) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS; HyClone Laboratories, Logan, UT, United States), a 1% penicillin-streptomycin solution, and 2 mM L-glutamine in a humidified incubator containing 5% CO2 at 37°C.
10
Murine Tumor Xenograft Modeling
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Murine B16F10 melanoma cells, MC38 colon adenocarcinoma, and EL4 thymoma cell lines were purchased from the American Type Culture Collection (ATCC) and kept as frozen stock in 2015. These cell lines have not been authenticated by the laboratory. Cells were cultured as described by the company.
The back region of female animals (~8-weeks old) was shaved one day prior to subcutaneous (s.c.) injection if the indicated cell line in 100ul of PBS immediately after the cells were washed with ice-cold PBS. 40,000 of B16F10, 250,000 MC38, or 200,000 EL4 cells were injected per mouse, unless otherwise indicated. Tumor progression was quantified using the formula V = (L*W2)/2, where V is volume, L is maximum length, and W is width perpendicular to the length. Excised tumors were weighed using a balance.
The back region of female animals (~8-weeks old) was shaved one day prior to subcutaneous (s.c.) injection if the indicated cell line in 100ul of PBS immediately after the cells were washed with ice-cold PBS. 40,000 of B16F10, 250,000 MC38, or 200,000 EL4 cells were injected per mouse, unless otherwise indicated. Tumor progression was quantified using the formula V = (L*W2)/2, where V is volume, L is maximum length, and W is width perpendicular to the length. Excised tumors were weighed using a balance.
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