Intracellular Staining by Flow Cytometry
|Detection of CCL2/JE/MCP‑1 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes treated with LPS for 48 hours were stained with Rat Anti-Mouse CCL2/JE/MCP‑1 APC-conjugated Monoclonal Antibody (Catalog # IC479A, filled histogram) or isotype control antibody (Catalog # IC013A, 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.|
Mouse CCL2 is a member of the beta (C-C) subfamily of chemokines. The mouse CCL2 gene was initially identified as a platelet-derived growth factor-inducible gene in mouse fibroblasts. Mouse CCL2 cDNA encodes a 148 amino acid (aa) residue with a putative 23 aa signal peptide that is cleaved to generate the mature protein. Mouse CCL2 shares 82% amino acid sequence identity with rat CCL2. Mouse CCL2 also shares 55% amino acid sequence identity with human MCP-1. Compared to human MCP-1, mouse CCL2 has a 49 aa residue extension at the carboxy-terminus. When a DNA sequence encoding the 125 aa residue of the mature CCL2 protein was expressed in E. coli at R&D Systems, the purified protein had the predicted N-terminus but a mass of 8525 Da. The truncation of most of the C-terminal extension could be due either to purification artifact or to post-translational modification. The truncated recombinant CCL2 has a potency similar to that of human MCP-1 in the monocyte chemotaxis assay. Mouse CCL2 has full activity on human cells while human MCP-1 has limited activity on mouse cells.
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