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Detection of Cellular Response to an in vitro Challenge with Bacterial Gram-Negative Lipopolysaccharides (LPS) in Peripheral Blood Mononuclear Cells (PBMCs)
Conclusion
The
in vitro
whole blood PBMC stimulation model combined with a simple
sample preparation workflow strategy allows for a facile and reproducible
method of testing cell signaling in the immune response from human research
samples.
Spectral Library creation from simple fractionation techniques allows for a
large number of different PTMs to be identified without using any specific PTM
enrichment strategy.
Pinpoint software allows for targeted quantification in even full scan discovery
modes in complex sample mixtures.
Future work will be to utilize the library and initial data to generate a targeted
list of peptides for selected reaction monitoring (SRM) workflows on both
Thermo Scientific™ TSQ Vantage™ and Thermo Scientific™ Q Exactive™
instruments. These will be to address the low level library peptides that were
difficult to quantify in this workflow, but are of interest in the biological
pathways that were shown to be up- and down-regulated in these samples and
are also correlated with sepsis.
References
1.
Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Critical Care Med 2001,
(29), 1303-1310
2.
Robbins Basic Pathology. 8th edition
3.
Mendes ME, Baggio-Zappia GL, Brunialti MK, Fernandes Mda L, Rapozo MM,
Salomao R. Immunobiology, 2011 Mar;216(3):285-95.
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This information is not intended to encourage use of these products in any manners that might infringe the
intellectual property rights of others.
For Research Use Only. Not for use in diagnostic procedures.
ted from LPS stimulation. Peptide
beta-1 precursor gi|63025222
uantifications from the first two
e peptide GDDTPLHLAASHGHR in
ts (pairs, left to right).
FIGURE 9. MyD88 signal pathway proteins up-regulated from LPS stimulation.
Peptide QILEGLK from mitogen-activated protein kinase kinase kinase 15
gi|282847398 [Homo sapiens].
te of peptide
otein kinase gi|4758606 [Homo
High Stimulation B) Alignment of
B