Monoclonal antibody- clone M7/1 recognizes human endothelial-monocyte-activating polypeptide II (EMAP II). EMAPII is a proinflammatory cytokine and chemoattractant of macrophages and polymorphonuclear lymphocytes. The protein was initially identified as a product of murine methylcholanthrene A-induced fibrosarcoma cells. EMAP II is synthesized as a 34 kDa precursor molecule (proEMAP),- it can run anomalously in SDSpage at 43 kDa, and is enzymatically cleaved to produce a biologically active 22 kDa mature polypeptide. It modulates a range of properties of endothelial cells, monocytes, and neutrophils in vitro, and induces an acute inflammatory reaction and tumor regression in vivo. Later it was found to be anti-angiogenic and capable of induction of apoptosis in proliferation and hypoxic endothelial cells in vitro in tumor vasculature in vivo. EMAP II is released from cells, by a yet unknown mechanism, as pro- and mature EMAP II proteins in response to various forms of cellular stresses, including glucose starvation and hypoxia. Endothelial cells are the main target- for EMAPII and explain the anti-angiogenic property. The anti-tumor effect is expected to a large extent to be mediated through this characteristic. EMAPII is also associated with brain diseases and injury. EMAPII expression is related with leukocytes-infiltrated inflamed tissue and subsequent apoptosis. In solid tumors there is also an immunosuppressive role identified. EMAPII inducing tumor cells induce apoptosis of lymphocytes. EMPAII is considered a marker for microglial cells and macrophages in brain diseases. The distribution of EMAPII and its precursor in biopsies might be applicable as additive diagnostic tool.
Flow cytometry, Functional studies, Immuno assays, Western blot
For immune assay and Western blotting, 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. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.