The mouse macrophages cell line RAW264.7 (ATCC) was grown in DMEM medium, supplemented with 10% heat-inactivated FBS (endotoxin-free Certified FBS; Invitrogen), 2% penicillin/streptomycin, 2 mM l -glutamine, and 10 mM HEPES (all from Gibco). The cells were seeded in T-75 tissue culture flasks (Falcon, USA) and used between passages 2 and 3. For quality control, we tested the cells for mycoplasma with the PlasmoTest kit (Invivogen). HDAC1 knockdown cells were generated by shRNA and validated by Western blotting analyses as described in our previous publication (20 (link)) and grown in a complete DMEM medium in the presence of puromycin (10 μg/mL). The primary antibody to detect HDAC1 was specific to HDAC1 (cat no. 5356; Cell Signaling Technology).
Endotoxin free certified fbs
Manufactured by Thermo Fisher Scientific
Endotoxin-free Certified FFS is a high-quality fetal bovine serum that has been screened and certified to be free of endotoxins. It is a critical component for cell culture applications that require a low-endotoxin environment.
Automatically generated - may contain errors
Lab products found in correlation
T 75 tissue culture flasks, by BD (3 mentions)
Hepes, by Thermo Fisher Scientific (3 mentions)
Plasmotest kit, by InvivoGen (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Thp1 blue cells, by InvivoGen (2 mentions)
Nf κb inducible reporter, by InvivoGen (2 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Raw264, by American Type Culture Collection (1 mentions)
3 protocols using endotoxin free certified fbs
1
Generation and Validation of HDAC1 Knockdown Cell Line
2
NF-κB Activation in Monocytic Cell Lines
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All cells were maintained in 5% CO2 at 37°C. THP-1 Blue cells (human monocytic leukemia line with an NF-κB–inducible reporter (Invivogen) and RAW264.7 cells carrying the NF-κB luciferase plasmid (IMGENEX, USA) were maintained in RPMI 1640 medium (Invitrogen) and Iscove's modified Dulbecco's medium (IMDM, Invitrogen), respectively. The media were supplemented with 10% heat-inactivated FBS (endotoxin-free Certified FBS; Invitrogen), 2% penicillin/streptomycin, 2 mM L-glutamine, and 10 mM HEPES (all from Gibco). The cells were seeded in T-75 tissue culture flasks (Falcon, USA) and used between passages 2 and 3.The THP-1 and RAW264.7 cell lines were extensively validated by the Invivogen and IMAGENEX respectively. For quality control, we have tested the cells for mycoplasma with PlasmoTest™ kit (Invivogen).
3
NF-κB Activation in Monocytic Cell Lines
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