Detection of S100A8/S100A9 Heterodimer in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P) and either (A) Mouse Anti-Human S100A8/S100A9 Heterodimer Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC9337G) or (B) Mouse IgG2A Alexa Fluor 488 Isotype Control (Catalog # IC003G). 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.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
Background: S100A8/S100A9 Heterodimer
S100A8 (also known as MRP8, Calgranulin A, and CP-10) and S100A9 (also known as MRP14 and Calgranulin B) are pro-inflammatory members of the S100 family of secreted calcium binding proteins (1, 2). They are up-regulated in neutrophils and monocytes at sites of inflammation (e.g. psoriasis, rheumatoid arthritis, cardiac ischemia) and are present at elevated concentrations in rheumatoid arthritis synovial fluid (3-5). The 10 kDa human S100A8 and 14 kDa S100A9 each contain two EF‑hand calcium binding motifs. Human S100A8 shares 57% and 61% amino acid (aa) sequence identity with mouse and rat S100A8, respectively. Human S100A9 shares 57% and 62% amino acid sequence identity with mouse and rat S100A9, respectively (6, 7). In the presence of calcium or zinc, S100A8 and S100A9 associate into non-covalent homodimers and 34-35 kDa heterodimers with each other (8-10). The heterodimer additionally binds and sequesters manganese, thereby restricting the growth of Mn-dependent bacteria (11). The S100A8/A9 heterodimer exhibits functions beyond those performed by the individual proteins. These include binding to fatty acids such as arachidonic acid and promoting astrocyte proliferation (3, 12). S100A8, S100A9, and the heterodimer each promote neutrophil infiltration into sites of inflammation and inflammatory cytokine production by monocytes (4, 5, 9).
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S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9
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