Subcellular Fractionation Studies on Macrophages

Kdo2-Lipid A Experiments

Experiments were conducted with RAW264.7 cells fed 10% fetal calf serum, as per protocol. Kdo2-Lipid A-treated and control cells were harvested at 24 hrs post treatment and fractionated using combination of differential centrifugation and isopycnic gradients using protocol PP0000004301(pdf). Three separate experiments (biological replicates) were carried out, and the data presented are mean±SE.
It should be noted that lipid measurements for each subcellular fraction are expressed as pmol/mg protein. Because the amounts of each fraction within a cell and also the recovery efficiencies are different, one cannot assume that the pmol values for a particular lipid represent the total amounts in each subcellular organelle per cell.

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Averages of biological replicates, by analyte category

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References

Subcellular organelle lipidomics in TLR-4-activated macrophages Andreyev AY, Fahy E, Guan Z, Kelly S, Li X, McDonald JG, Milne S, Myers D, Park H, Ryan A, Thompson BM, Wang E, Zhao Y, Brown HA, Merrill AH, Raetz CR, Russell DW, Subramaniam S, Dennis EA. J Lipid Res 51, 2785-97 (2010)
PMID: 20574076

Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4. Raetz C, Garrett T, Reynolds C, Shaw W, Moore J, Smith JD, Ribeiro A, Murphy R, Ulevitch R, Fearns C, Reichart D, Glass C, Benner C, Subramaniam S, Harkewicz R, Bowers-Gentry R, Buczynski M, Cooper J, Deems R and Dennis E.Journal of Lipid Research 47, 1097-1111 (2006)
PMID: 16479018 [doi:10.1194/jlr.M600027-JLR200]

logo LIPID MAPS is funded by a Wellcome Trust.