Anti-Mouse Siglec-F-BX016; Alexa Fluor™ 488 - RX016 (C2878-25Tests)

Siglecs (sialic acid-binding immunoglobulin superfamily lectins) are a family of single pass, transmembrane cell surface proteins characterized by shared structural motifs and an ability to recognize sialic acids1,2. Siglec-F selectively recognizes 6'-sulfo-sialyl Lewis X as a glycan ligand, and this interaction is inhibited by the E50-2440 antibody3. Siglec-F belongs to the Siglec-3 (CD33) group4 and is a functionally convergent paralog of human Siglec-83. Mouse Siglec-F is used as a model to understand glycan based therapeutic targeting strategies in humans1, 2. Expression of Siglec-F on eosinophils3 plays a role in the allergic inflammatory response, during which its surface levels on eosinophils and other cells increase1. Siglec-F deficient mice that are exposed to allergen sensitization and asthma-inducing conditions display more pronounced bone marrow, blood, and tissue eosinophilia due to reduced apoptosis1 as well as exaggerated eosinophilic inflammation and tissue remodeling in asthma models2. Administration of Siglec-F antibody reduces blood and tissue eosinophils in vivo2. The rat monoclonal antibody E50-2440 was prepared by immunizing Lou rats with mSiglec-F-Fc, a fusion protein of the first two Ig-like domains of mouse Siglec-F and human IgG Fc fragment separated by an enterokinase cleavage site/FLAG tag (DYKDDDDK)4. Spleen cells were fused with Yb/20 myeloma cells, and hybridomas were screened by ELISA, tested by flow cytometry, and isotyped. The E50-2440 clone was selected based on its high staining intensity on primary cells. Siglec-F is a reliable marker of eosinophils among granulocytes in mouse1, and cellular staining profiles are identical in the C3H, Balb/c, and C57BL/6 mouse strains4.