The monoclonal antibody 27E10 recognizes an epitope specific for the S100A8/A9 heterocomplex that is not exposed on the individual subunits S100A8 (MRP8, calgranulin-A) or S100A9 (MRP14, calgranulin-B). The calcium-binding migration inhibitory factor-related proteins, MRP-8 (S100A8) and MRP-14 (S100A9) belong to the S100 protein family. The expression of these proteins is largely confined to the cytosol of neutrophils and monocytes. The complex formation of these proteins is a calcium-dependent process. The S100A8/A9 heterocomplex, also called MRP-8/MRP-14 complex or calprotectin, comprises 60% of the cytoplasmic protein fraction of circulating polymorphonuclear granulocytes and is also found in monocytes, macrophages and ileal tissue eosinophils. Peripheral blood monocytes carry the antigen extra- and intracellularly, neutrophils only intracellularly. The S100A8/A9 complex has antibacterial, antifungal, immunomodulating and antiproliferative effects. Besides this it is a potent chemotactic factor for neutrophils. Plasma concentrations are elevated in diseases associated with increased neutrophil activity, like inflammatory bowel disease. Granulocytes terminate their existence after transmigration through the intestinal wall. Therefore calprotectin is also detectable in feces. Elevated levels of calprotectin have been observed in body fluids such as plasma, saliva, gingival crevicular fluid, stools, and synovial fluid during infection and inflammatory conditions.
The monoclonal antibody 27E10 can be used for early detection of inflammatory macrophages, for the characterization of tumorous tissues and the monitoring of peripheral blood cell cultures. The antibody 27E10 does not react with lymphocytes or platelets.
Flow cytometry, Frozen sections, Immuno assays, Immuno fluorescence, Immuno precipitation, Western blot
W: non-reduced;~22 kDa; doesn’t recognize the single proteins S100A8 and S100A9
HPLC: reduced (~17 kDa) and non-reduced(various sizes due to association with other elements (Ref1)
F: acetone fixation; 0.1 % hydrogen peroxide treatment to reduce enogenous peroxidise activity; positive control: inflammatory tissue; negative control: normal human tissue (skin, lung, colon) (Ref1)
FC: Extracelullar expression on monocytes, as negative control HL-60, platelets, lymphocytes can be used.
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Platelets, lymphocytes, HL-60 cells