C2C12 cells were used for co-immunoprecipitation and ChIP experiments, COS-7 cells were used for luciferase assays, and Sf9 cells were used to produce PAX7 protein for EMSAs. C2C12, COS-7, and Sf9 cells were purchased from and authenticated by ATCC. These cells were not contaminated by mycoplasma as determined by the MycoSensor PCR Assay Kit (Agilent Technologies). C2C12 and COS-7 cells were cultured at 37 °C in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum, and 1% penicillin/streptomycin, while Sf9 cells were cultured in SF-900 III SFM media (Life Technologies).
Sf 900 3 sfm media
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Sf-900 III SFM media is a serum-free, insect cell culture medium designed for the growth and maintenance of insect cells. It provides the necessary nutrients and growth factors to support insect cell lines, such as Sf9 and Sf21, in suspension culture. The Sf-900 III SFM media is formulated to support high-density cell growth and high-level protein expression.
Automatically generated - may contain errors
Lab products found in correlation
Sf9 cells, by Thermo Fisher Scientific (3 mentions)
Freestyle 293 expression medium, by Thermo Fisher Scientific (2 mentions)
Hek293s gnti cells, by American Type Culture Collection (2 mentions)
Pen strep, by Thermo Fisher Scientific (2 mentions)
Freestyle 293 media, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
Mycosensor pcr assay kit, by Agilent Technologies (1 mentions)
Express five sfm, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Freestyle 293 medium, by Thermo Fisher Scientific (1 mentions)
Fugene reagent, by Promega (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Countess cell counter, by Thermo Fisher Scientific (1 mentions)
Recombinant human serum albumin, by Merck Group (1 mentions)
Bacpak baculovirus rapid titer kit, by Takara Bio (1 mentions)
Transit insect transfection reagent, by Mirus Bio (1 mentions)
11 protocols using sf 900 3 sfm media
1
Cell Lines for Protein Studies
2
Recombinant Tubulin Production in Sf9 Cells
3
Sf-9 and HEK293T Cell Culture for Protein Expression
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Sf-9 insect cells were cultured in Sf-900 III SFM media (Gibco, Waltham, Massachusetts, USA) at 27°C (without CO2 and humidification) on an orbital shaker platform and were used to generate baculoviruses and protein expression. HEK293T cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS), penicillin, and streptomycin at 37°C and 5% CO2. HEK293T cells in antibiotic-free media in a 15-cm dish were transfected with Strep-SMCR8 and C9orf72 plasmids using Lipofectamine 2000 and were grown for 48 hours post-transfection. Moreover, 90 minutes prior to the lysis, cells were starved using amino acid- and glucose-free media.
4
Rat GluK2 Expression in HEK Cells
5
Heterologous Expression of Glutamate Receptors
6
Insect Cell Lines for Baculovirus and Protein Production
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Insect cells used for the baculovirus preparation were Spodoptera frugiperda 21 (Sf21) (source EMBL), grown in Sf-900™ III SFM media (Gibco, 12658027), as published previously (Roostalu et al., 2018 (link)). Insect cells used for recombinant protein expression were Trichoplusia ni High Five insect cells, grown in Express Five™ SFM (Gibco, 10486025).
7
TRPM7 Expression in HEK293 Cells
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For structural experiments, expression of mouse TRPM7 was performed in HEK293S cells lacking N-acetyl-glucosaminyltransferase I (GnTI−, mycoplasma test negative, ATCC #CRL-3022) that were maintained at 37°C and 6% CO2 in Freestyle 293 medium (Thermo Fisher Scientific #12338-018) supplemented with 2% FBS. Baculovirus for infecting HEK293S GnTI− cells was produced in Sf9 cells (GIBCO) cultured in the Sf-900 III SFM media (GIBCO) at 27°C. For aequorin-based Ca2+ influx assay and patch-clamp experiments, TRPM channels were expressed in HEK293T cells (mycoplasma test negative, ATCC #CRL3216) that were maintained at 37°C and 5% CO2 in DMEM (Merck, #D6429) supplemented with 10% fetal calf serum (Thermo Fisher Scientific, #10270106), 100 μg/ml streptomycin and 100 U/ml penicillin (Merck, #P4333).
8
Recombinant Protein Expression in E. coli and Baculovirus
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E. coli cells of BL21-CodonPlus(DE3)-RIL strain transformed with expression plasmids were cultured in 2xYT media (16 g/L tryptone, 10 g/L yeast extract, 5 g/L NaCl) supplemented with ampicillin and chloramphenicol at 37°C. Protein expression was induced by adding IPTG to the final concentration of 0.2 mM when OD600 of the culture reached 0.6 and further incubation at 20°C for 16 hr.
Plasmids of pBIG1 derivatives carrying expression cassettes were recombined to baculoviral genome by Tn7 transposition in E. coli DH10EMBacY cells (Trowitzsch et al., 2010 (link)). Recombined EMBacY with expression cassettes were extracted from bacteria cells and used to transfect Sf9 cells for baculovirus generation. Initial viruses were obtained after 3 days incubation of EMBacY and Sf9 cells with FuGENE reagent (Promega). Viruses were amplified using Sf9 cells. Litter-scale protein expression was performed by incubation of viruses and Tnao38 cells at 27°C for 4 days with starting density of 1.2 million Tnao38 cells per mL of Sf-900 III SFM media (Thermo Fisher Scientific).
Plasmids of pBIG1 derivatives carrying expression cassettes were recombined to baculoviral genome by Tn7 transposition in E. coli DH10EMBacY cells (Trowitzsch et al., 2010 (link)). Recombined EMBacY with expression cassettes were extracted from bacteria cells and used to transfect Sf9 cells for baculovirus generation. Initial viruses were obtained after 3 days incubation of EMBacY and Sf9 cells with FuGENE reagent (Promega). Viruses were amplified using Sf9 cells. Litter-scale protein expression was performed by incubation of viruses and Tnao38 cells at 27°C for 4 days with starting density of 1.2 million Tnao38 cells per mL of Sf-900 III SFM media (Thermo Fisher Scientific).
9
Baculoviral and Mammalian Cell Culture Protocols
10
Baculovirus and HEK293S Protein Expression
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