Human immortalized keratinocytes (HaCaT) were purchased from ATCC (Manassas, VA, USA) and maintained according to the manufacturer's instructions. Briefly, keratinocytes were cultured in DMEM medium (Corning, Manassas, VA) supplemented with 10% bovine calf serum (HyCloneTM Laboratories, Logan, UT). HOCl exposure was performed in PBS in order to minimize the impact of indirect chlorination reactions (e.g. HOCl-modification of serum proteins in medium that might confound cellular effects). Cells were maintained in a humidified incubator (37 °C, 5% CO2 and 95% air) [36 (link),37 (link)].
Bovine calf serum (bcs)
Manufactured by Corning
Bovine calf serum is a common cell culture supplement derived from the blood of bovine calves. It provides a complex mixture of nutrients, growth factors, and other components essential for the growth and maintenance of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Hacat, by American Type Culture Collection (1 mentions)
Dmem medium, by Corning (1 mentions)
Glass bottom imaging dishes, by MatTek (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Corning (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
2 protocols using bovine calf serum (bcs)
1
HOCl Exposure on Immortalized Keratinocytes
2
NIH 3T3 Fibroblast Cell Culture Protocol
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NIH 3T3 mouse fibroblast cells
were grown in DMEM (Corning) supplemented with 10% bovine calf serum
(Corning) and 1% penicillin/streptomycin (Corning) at 37 °C in
a 5% CO2 atmosphere. For serum starvation, NIH 3T3 cells
were cultured in DMEM supplemented with 1% penicillin/streptomycin
(serum-free medium). For experiments, cells were seeded in 12-well
plates for Western blot analysis, 35 mm glass-bottom imaging dishes
(MatTek Corporation) for immunofluorescence, and 35 mm dishes for
HPLC and LC–MS analysis. For C1-EGFP translocation assays,
cells seeded on 35 mm imaging dishes were transfected using 1 μg
of C1-EGFP plasmid (a gift from the De Camilli laboratory, Yale University,
New Haven, CT) and 3 μL of Lipofectamine 2000 (Invitrogen) per
dish, 24 h prior to the experiment.
were grown in DMEM (Corning) supplemented with 10% bovine calf serum
(Corning) and 1% penicillin/streptomycin (Corning) at 37 °C in
a 5% CO2 atmosphere. For serum starvation, NIH 3T3 cells
were cultured in DMEM supplemented with 1% penicillin/streptomycin
(serum-free medium). For experiments, cells were seeded in 12-well
plates for Western blot analysis, 35 mm glass-bottom imaging dishes
(MatTek Corporation) for immunofluorescence, and 35 mm dishes for
HPLC and LC–MS analysis. For C1-EGFP translocation assays,
cells seeded on 35 mm imaging dishes were transfected using 1 μg
of C1-EGFP plasmid (a gift from the De Camilli laboratory, Yale University,
New Haven, CT) and 3 μL of Lipofectamine 2000 (Invitrogen) per
dish, 24 h prior to the experiment.
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