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Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling by array of mouse intestinal epithelial m-ICcl2 cells stimulated with lipopolysaccharide

ABSTRACT: m-ICcl2 cells (Bens et al., Am. J. Physiol., 1996) were grown for 6 days on collagen coated cell culture flasks in medium as recommended (see ference above). Medium was changed every 48 hours. The cells were mycoplasma tested (PCR) and found negative. The medium and PBS was endotoxin tested (ELISA) and found to be negative (< 50pg/mL). Unstimulated cells (control) were left cultured in medium only. Lipopolysaccharide (100 ng/mL in water, D31m4 E. coli LPS from List Biochemicals) was sonicated (10 min_ prior to use. Cells were stimulated 6 hours with 100 ng/mL LPS and harvested or stimulated with LPS (100 ng/mL) and washed with warm phosphate buffered saline (PBS) after 6 hours, followed by medium change every 48 hours. Cells were harvested after 96 hours (6 hours incubated in the presence of LPS, 90 hours after stimulation). RNA was isolated after repeated (3x) washing with PBS using Trizol reagent (1 mL/75 cm2) using the standard protocol and resuspended in Millipore water. Quantification was done by spectroscopy and the RNA was stored at -80C.

ORGANISM(S): Mus Musculus

SUBMITTER: Mathias Hornef

PROVIDER: E-MTAB-355 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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