Technical Data
N2254
Neutrophil
Description:
The Ly-6G/-6C locus encodes a family of Ly-6 proteins. Ly-6 antigens have a molecular weight between 15,000-18,000. Ly6G is together with Ly6c a component of the myeloid differentiation antigen Gr-1. Ly6G, a GPI-anchored protein, is a good marker of peripheral neutrophils. Although predominantly present on neutrophils, it is also expressed on a subset of eosinophils, differentiating premonocytes and plasmacytoid dendritic cells. Ly6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon gamma. It 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. The physiological role of Ly6G alone remains unclear. N2254 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 an inhibitory effect on local immune responses. It has been shown to be useful for depletion of neutrophils in mice, depleting neutrophils as soon as 6hr after injection and up to 6 days.

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

Recommended Dilution:
Flow Cytometry: 10ug/ml labeled 5x10e5 cells.
Immunohistochemistry (Frozen): 1:50 Acetone fixation
Immunohistochemistry (paraffin): 1:50 Block with 20% normal rabbit serum.
Functional Studies: Neutrophil depletion. Mice treated with N2254 given intraperitoneally at a dose of 1mg, 6 hours
before infection.
Optimal dilutions to be determined by the researcher.

Positive Control: Mouse neutophils
Negative Control: Mouse Thymocytes

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG2b6A608Affinity Purified
SizeStorageShippingSourceHost
100ug-20CBlue IceMouseRat
Concentration:
~0.1mg/ml
Immunogen:
Purified BALB/c mouse neutrophils.
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in PBS, 0.1% BSA.
Specificity:
Recognizes mouse Neutrophil. Highly specific for mouse Ly-6G and Ly-6C.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Lopez, A et al; Brit J Haematology (1984) 57: 489. 2. McLaren, D et al; Parasite Immunol (1987) 9: 667. 3. Van Lent, P et al; J Leukoc Biol (1997) 61: 267. 4. Lubberts, E et al; J Immunol (1999) 163: 4546. 5. Tacchini-Cottier, F et al; J Immunol (2000) 165: 628. 6. Vries de, B et al; J Immunol (2003) 170: 3883. 7. Nagendra, S et al; Cytometry A (2004) 58: 195. 8. Navarini, A et al; PNAS (2009) 106: 7107.