Intracellular Staining by Flow Cytometry
|Detection of CCL3/MIP‑1 alpha in Human Blood Monocytes by Flow Cytometry. Human peripheral blood monocytes treated with LPS were stained with Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P) and either (A) Mouse Anti-Human CCL3/MIP‑1 alpha APC‑conjugated Monoclonal Antibody (Catalog # IC2701A) or (B) Mouse IgG2B Allophycocyanin Isotype Control (Catalog # IC0041A). 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.|
The macrophage inflammatory proteins -1 alpha and -1 beta were originally co-purified from medium conditioned by an LPS-stimulated murine macrophage cell line. Human MIP-1 alpha refers to the products of several independently cloned cDNAs, including LD78, pL78, pAT464, and GOS19. These cDNAs all code for the same human protein that is a homologue of the murine MIP-1 alpha. Mature MIP-1 alpha and MIP-1 beta in both human and mouse share approximately 70% homology at the amino acid level. The MIP‑1 proteins are members of the beta (C-C) subfamily of chemokines.
Both MIP-1 alpha and MIP-1 beta are monocyte chemoattractants in vitro. Additionally, the MIP-1 proteins have been reported to have chemoattractant and adhesive effects on lymphocytes, with MIP-1 alpha and MIP-1 beta preferentially attracting CD8+ and CD4+ T cells, respectively. MIP-1 alpha has also been shown to attract B cells as well as eosinophils. MIP-1 proteins have been reported to have multiple effects on hematopoietic precursor cells and MIP-1 alpha has been identified as a stem cell inhibitory factor that can inhibit the proliferation of hematopoietic stem cells in vitro as well as in vivo. The functional receptor for MIP-1 alpha has been identified as CCR1 and CCR5.
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