Description
The 6F12 antibody reacts with a 7-transmembrane-domain protein, which is similar to the F4/80 macrophage antigen of the EGF-TM7 protein family and is encoded by the Emr4 gene. The FIRE protein is expressed on myeloid cells with a denditic cell (DC) developmental potential, including subsets of DC and macrophages in the spleen and lymph nodes, most resident peritoneal macrophages, many peripheral blood monocytes, and a subpopulation of bone-marrow myeloid-cell progenitors. The protein is not detected on peripheral T and B lymphocytes, and it is down-regulated on thioglycollate-elicited peritoneal macrophages and on dendritic cells activated by GM-CSF, IFN-γ, anti-CD40, and LPS. Using soluble biotinylated fusion protein, a FIRE ligand was detected on a mouse IgG+ B lymphoma cell line (A20), but not on myeloid, fibroblast, or T-cell lines, suggesting that the FIRE protein may be involved in immunoregulatory interactions between antigen-presenting cells and B lymphocytes.
For flow cytometry of leukocyte suspensions, we recommend that Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) be used. Since this antigen is expressed at low density on myeloid cells, it may be desirable to amplify staining by using a biotinylated second-step antibody followed by a "bright" third-step reagent, such as Streptavidin-APC (Cat. No. 554067). If Mouse BD Fc Block™ antibody is used, it is important that the second-step anti-rat IgG does not react with 2.4G2 mAb (rat IgG2b, κ); we recommend biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894).
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
