HEK293F cells were grown in Dulbecco’s modified eagle medium (Capricorn Scientific, Ebsdorfergrund, Germany) supplemented with 10% fetal calf serum and 1% penicillin/streptomycin (PAN-Biotech, Aidenbach, Germany) and were transfected with β3 (wild-type or mutant) together with full-length wild-type αIIb or αv in pcDNA4/HisMax©-TOPO® Zeocin (Invitrogen) vector using jetOptimus Transfection reagent (Polyplus, Illkirch, France). After 4 weeks of cell culture in the presence of Geneticin (400 μg/mL) and Zeocin (200 μg/mL), cell lines were analyzed for αIIbβ3 and αvβ3 surface expression by flow cytometry. Stable transfectants expressing high αIIbβ3 and αvβ3 densities on the cell surface were sorted with Alexa Flour 488-labeled mAb Gi5 (anti-αIIbβ3 complex) using a BD FACSAria III with FACSDiva 6.1.3 software (Becton and Dickinson).
Dulbecco s modified eagle medium
Manufactured by Capricorn
Sourced in Germany
Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium that provides nutrients and growth factors essential for the maintenance and proliferation of various cell types. It is a widely used basal medium in cell biology and tissue engineering applications.
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Lab products found in correlation
Fetal bovine serum (fbs), by Capricorn (2 mentions)
Facsaria 3, by BD (1 mentions)
Jetoptimus transfection reagent, by Polyplus Transfection (1 mentions)
Penicillin, by Capricorn (1 mentions)
Streptomycin, by Capricorn (1 mentions)
L glutamine, by Capricorn (1 mentions)
Sodium pyruvate, by Capricorn (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicilin, by Lonza (1 mentions)
Galaxy 170s humidified incubator, by Eppendorf (1 mentions)
Streptomycin, by Lonza (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Dulbecco modified eagle medium (dmem), by Capricorn (1 mentions)
Trypsin edta, by Capricorn (1 mentions)
Stable glutamine, by Capricorn (1 mentions)
Antibiotic antimycotic solution, by Capricorn (1 mentions)
5 protocols using dulbecco s modified eagle medium
1
Generation of HEK293F Stable Cell Lines Expressing αIIbβ3 and αvβ3 Integrins
2
Establishment of BRAFV600E-Mutant Cell Lines
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Human colon carcinoma cell line RKO and human melanoma cell line A375 both harbouring the BRAFV600E mutation were purchased from the American Type Culture Collection (Manassas, VA, USA). Both RKO and vemurafenib-resistant RKO cells were maintained in Eagle’s Minimum Essential Medium (Capricorn Scientific, Ebsdorfergrund, Germany) supplemented with 10% fetal bovine serum (Capricorn Scientific, Germany), 2 mM L-glutamine (Capricorn Scientific, Germany), penicillin (100 U/mL) (Capricorn Scientific, Germany) and streptomycin (100 µg/mL) (Capricorn Scientific, Germany) in humified atmosphere with 5% CO2 at 37 °C. Melanoma cell line A375 and its vemurafenib-resistant counterpart were maintained in Dulbecco’s Modified Eagle Medium (Capricorn Scientific, Germany) supplemented with 10% fetal bovine serum (Capricorn Scientific, Germany), 2 mM L-glutamine (Capricorn Scientific, Germany), penicillin (100 U/mL) (Capricorn Scientific, Germany), streptomycin (100 µg/mL) (Capricorn Scientific, Germany) and 1 mM sodium pyruvate (Capricorn Scientific, Germany) in humified atmosphere with 5% CO2 at 37 °C.
3
Cell Culture in Supplemented DMEM
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Cells were maintained in Dulbecco’s Modified Eagle Medium (containing 4.5 g/L d-glucose, stable glutamine, sodium pyruvate; Capricorn Scientific) supplemented with 10% Fetal Bovine Serum (Sigma-Aldrich), 100 IU/mL penicilin and 100 mg/mL streptomycin (Lonza). Cells were grown at 37°C in a Galaxy 170s humidified incubator (Eppendorf) with a 5% CO2 atmosphere.
4
HEK293T Cell Culture Protocol
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HEK293T cells (ATCC) were cultured in Dulbecco’s modified Eagle medium (DMEM; Capricorn scientific) supplemented with 10% fetal bovine serum (FBS; Corning), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco). Cells were maintained at 37 °C and in a humidified, 5% CO2 atmosphere.
5
Lycopene Cytotoxicity Evaluation
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Dulbecco's Modified Eagle Medium (DMEM), Fetal Bovine Serum (FBS), stable glutamine, antibiotic-antimycotic solution, trypsin-EDTA and Hank's Balanced Salt Solution (HBSS) were purchased from Capricorn (Germany).Lycopene (Lyc) and Neutral red (NR) were purchased from Sigma-Aldrich, while (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) was obtained from Carl Roth (Germany). Methotrexate (MTX), (Metoject,10 mg/ml) injection solution was purchased from Medac GmbH (Germany).
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