The Ly-6G/-6C locus encodes a family of Ly-6 proteins including Ly-6G and Ly-6C. Ly-6 antigens have a molecular weight between 15,000 and 18,000. Ly6G is together with Ly6c a component of the myeloid differentiation antigen Gr-1. Ly6G a GPI-anchored protein and is a good marker of peripheral neutrophils. Although predominantly presents on neutrophils, it is also expressed on a subset of eosinophils, differentiating pre- monocytes and plasmacytoid dendritic cells. Ly6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon gamma, and may play a role in the development and maturation of lymphocytes. It is expressed on bone marrow cells, monocytes/macrophages, neutrophils, endothelial cells, and T cell subsets. Expression of Gr-1 in bone marrow correlates with granulocyte differentiation and maturation. However, the physiological role of Ly6G alone remains still unclear. The monoclonal antibody NIMP-R14 has been successfully used to stain polymorphonuclear (PMN) cells and monocytes for fluorescent activated cell sorting and in frozen and paraffin sections. Treatment with antibodies in vivo leads to neutropenia and has inhibitory effect on local immune responses. Furthermore, it has been shown to be useful for depletion of neutrophils in mice. It depletes neutrophils as soon as 6 hours after injection and up to 6 days.

Applications:
Suitable for use in Flow Cytometry, FLISA, Functional Studies and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunohistochemistry (frozen): fixed with acetone
Immunohistochemistry (Paraffin): Blocking with 20% normal rabbit serum
Flow Cytometry: 10ul/250.000 cells in a 100ul total staining volume
Functional Studies: Neutrophil depletion. Mice were treated with NIMP-R14 given intraperitoneally at a dose of 1mg, 6h before infection
Optimal dilutions to be determined by the researcher.

Positive Control: Mouse neutrophils

Negative Control: Mouse thymocytes

Storage and Stability:
Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: PE conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.