Intracellular Staining by Flow Cytometry
|Detection of TNF-alpha in EL‑4 Mouse Cell Line by Flow Cytometry. EL‑4 mouse lymphoblast cell line treated with anti-CD3/anti-CD28, PMA, and Calcium Ionomycin were stained with Rat Anti-Mouse TNF-alpha Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC410F, filled histogram) or isotype control antibody (Catalog # IC005F, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
TNF-alpha is a trimeric glycoprotein active in both membrane bound and secreted forms. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. TNF-alpha binds to TNF RI and TNF RII present on virtually all cell types, through which it triggers the activation of multiple signal transduction pathways and modulates the expression of a wide variety of genes.
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