Intracellular Staining by Flow Cytometry
|Detection of IL‑6 in J774A.1 Mouse Cell Line by Flow Cytometry. J774A.1 mouse reticulum cell sarcoma macrophage cell line treated with LPS was stained with Rat Anti-Mouse IL‑6 Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC406F, 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.|
Interleukin 6 (IL-6) is a pleiotropic alpha -helical cytokine that plays important roles in acute phase reactions, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6 activity is central to the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. It is secreted by multiple cell types as a 22 kDa-28 kDa phosphorylated and variably glycosylated molecule (1-4). Mature mouse IL-6 is 187 amino acids (aa) in length and shares 42% and 85% aa sequence identity with human and rat IL-6, respectively (5). Alternate splicing generates several isoforms with internal deletions (6). Mouse IL-6 is equally active on rat cells (7). IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R, triggering IL-6 R association with gp130 and gp130 dimerization (8). gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM (9). Soluble forms of IL-6 R are generated by both alternate splicing and proteolytic cleavage (9). In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R elicit responses from gp130-expressing cells that lack cell surface IL-6 R (3). Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous while that of IL-6 R is predominantly restricted to hepatocytes, leukocytes, and lymphocytes (3). Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R but not from other cytokines that utilize gp130 as a coreceptor (4, 10).
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.
The document you requested is not available online. Please enter the Catalog Number and Lot Number below to have a document emailed to you at the address provided