Intracellular Staining by Flow Cytometry
|Detection of IL‑1 beta /IL‑1F2 in Human Blood Monocytes by Flow Cytometry. Human peripheral blood monocytes either (A) untreated or (B) treated with 250 ng/mL LPS overnight were stained with Rabbit Anti-Human IL‑1 beta /IL‑1F2 APC‑conjugated Monoclonal Antibody (Catalog # IC8406A) and Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P). Quadrant markers were set based on control antibody staining (Catalog # IC105A). 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.|
IL-1 is a common name for two pleiotropic cytokines, IL-1 alpha (IL-1F1) and IL-1 beta (IL-1F2), which are the products of distinct genes. IL-1 alpha and IL-1 beta are structurally related polypeptides that share approximately 21% amino acid (aa) sequence identity in human. Both proteins are produced by a wide variety of cells in response to inflammatory agents, infections, or microbial endotoxins. While IL-1 alpha and IL-1 beta are regulated independently, they bind to the same receptor and may exert identical biological effects. IL-1 RI binds directly to IL-1 alpha or IL-1 beta and then associates with IL-1 R Accessory Protein (IL-1 R3/IL-1 R AcP) to form a high-affinity receptor complex that is competent for signal transduction. IL-1 RII has high affinity for IL-1 beta but functions as a decoy receptor and negative regulator of IL-1 beta activity. IL-1ra functions as a competitive antagonist by preventing IL-1 alpha and IL-1 beta from interacting with IL-1 RI (1-4). The human IL-1 beta cDNA encodes a 269 aa precursor that contains a 116 aa propeptide which is cleaved intracellularly by the cysteine protease IL-1 beta -converting Enzyme (Caspase-1/ICE) to generate an active cytokine (5-7). The 17 kDa mature human IL-1 beta shares 96% aa sequence identity with rhesus macaque, and 67-78% aa sequence identity with canine, cotton rat, equine, feline, mouse, porcine, and rat IL-1 beta.
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