Anti-Mouse CD16.2 (9E9) In Vivo Antibody- Low Endotoxin

According to surface plasmon resonance, 9E9 has strong reactivity to FcγRIV as well as low level binding to FcγRII and FcγRIII. In vivo, 9E9 binds and blocks FcγRIII only when 9E9 first binds FcγRIV on the same effector cell, resulting in concurrent inhibition of FcγRIII and FcγRIV. Native 9E9 binds to FcγRII and FcγRIII via the Fc. Blocking studies with 9E9 show that FcγRIV is necessary for IgG2a and IgG2b mediated platelet clearance in vivo. Additionally, blocking FcγRIV with 9E9 reduces B-cell depletion. 9E9 also interferes with immune complex binding to FcγRIV and can block FcγRIII on macrophages and neutrophils.

9E9 was produced by immunizing Armenian hamsters with an FcγRIV-IgG1 fusion protein consisting of the extracellular domain of FcγRIV fused to a mouse IgG1 Fc portion (D265A-variant deficient in Fc-receptor binding) . Splenic B cells were then fused to a mouse fusion partner, and hybridoma clones were screened for binding to CHO-K1-FcγRIV cells expressing FcγRIV.

This antibody is stable for at least one week when stored sterile at 2-8°C. For long term storage aseptically aliquot in working volumes without diluting and store at –80°C. Avoid Repeated Freeze Thaw Cycles.

FcγRIV is expressed on the cell membrane of splenic and bone marrow dendritic cells, monocytes, and macrophages as well as peripheral blood monocytes, neutrophils, thioglycollate-elicited macrophages, and myeloid cells. FcγRIV is absent from lymphoid populations, T cells, B cells, NK cells, and other granulocytes.