Using Image-Based Flow Cytometry with a FISH-Based FlowRNA Assay to Simultaneously Detect Intracellular TNF-α Protein and mRNA in Monocytes Following LPS Stimulation
| Autor: | Eric A. Prado, Adam S. Venable, Brian K. McFarlin, Andrea L. Henning |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Messenger RNA Lipopolysaccharide medicine.diagnostic_test Chemistry Monocyte CD14 Stimulation Molecular biology Flow cytometry 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology medicine.anatomical_structure medicine Tumor necrosis factor alpha Intracellular |
| Zdroj: | Imaging Flow Cytometry ISBN: 9781493933006 |
| Popis: | Existing methods of assessing monocyte inflammatory cytokine (IL-1β, IL-6, IL-8, and TNF-α) response to in vitro lipopolysaccharide (LPS) stimulation lack the ability to simultaneously detect intracellular mRNA and protein. This procedure takes advantage of new methodologies and instrumentation to simultaneously measure intracellular TNF-α mRNA and protein in CD14(+) monocytes after 1, 3, and 6 h of LPS stimulation. By assessing multiple timepoints, we are able to discern how LPS stimulation affects the temporal relationship between TNF-α mRNA and protein. By using image-based flow cytometry it is possible to co-localize mRNA and protein signals to identify the length of incubation that is needed to initiate protein translation. |
| Databáze: | OpenAIRE |
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