Intracellular Staining by Flow Cytometry
|Detection of CCL20/MIP‑3 alpha in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes treated with 1 μg/mL LPS for 48 hours were stained with Rat Anti-Mouse CCL20/MIP‑3 alpha Alexa Fluor® 700‑conjugated Monoclonal Antibody (Catalog # IC760N, filled histogram) or isotype control antibody (Catalog # IC005N, 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.|
CCL20, also known as MIP-3 alpha, LARC, and Exodus, is a beta chemokine. Mouse CCL20 cDNA encodes a 97 amino acid (aa) precursor protein with a 27 aa putative signal peptide that is predicted to be cleaved to form the 70 aa mature secreted protein. CCL20 is distantly related to other beta chemokines (20‑28% aa sequence identity). Mouse CCL20 shares approximately 71 and 63% amino acid sequence homology with rat and human CCL20, respectively. CCL20 has been shown to be expressed predominantly in lymph nodes, appendix, PBL, fetal liver, fetal lung, and epithelial cells of intestinal tissues. The expression of CCL20 is strongly upregulated by inflammatory signals and downregulated by the anti-inflammatory cytokine IL-10. CCL20 has been shown to be chemotactic for lymphocytes and dendritic cells, and inhibits proliferation of myeloid progenitors in colony formation assays. CCL20 is a functional ligand for CCR6 (previously referred to as GPR-CY4, CKR-L3, or STRL22 orphan receptor), a chemokine receptor that is selectively and highly expressed in human dendritic cells derived from CD34+ cord blood precursors.
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