Intracellular Staining by Flow Cytometry
|Detection of IL‑34 in NS0 Mouse Cell Line Transfected with Human IL-34 by Flow Cytometry. NS0 mouse myeloma cell line transfected with human IL-34 was stained with Mouse Anti-Human IL‑34 PE‑conjugated Monoclonal Antibody (Catalog # IC5265P, filled histogram) or isotype control antibody (Catalog # IC002P, 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 34 (IL-34; also known as uncharacterized protein C16orf77) is secreted as a homodimer consisting of 39 kDa monomers (1). It belongs to no known cytokine family. Human IL-34 is synthesized as a 242 amino acid (aa) precursor that contains a 20 aa signal sequence and a 222 aa mature chain. The mature chain contains one potential site of N-linked glycosylation. Human IL-34 is 71% identical to mouse IL-34 on the amino acid level (1). IL-34 is expressed in various tissues, including the heart, brain, liver, kidney, spleen, thymus, testes, ovary, small intestine, prostate, and colon, and is most abundant in the spleen (1). The receptor for IL-34 is colony-stimulating factor 1 receptor (CSF-1R) (1,2). IL-34 stimulates monocyte proliferation (1). In functional studies, IL-34, like CSF-1, the other ligand for CSF-1R, stimulated phosphorylation of extracellular signal-regulated kinase-1 and -2 (ERK1/2) in human monocytes (1). In addition, IL-34 promoted the formation of the colony-forming unit-macrophage (CFU-M), a macrophage progenitor, in human bone marrow cultures (1,3).
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