Dectin 2a, Recombinant, Mouse (CLEC6A, CLECSF10, NKCL)

Dectin-2alpha, also known as CLEC6A, CLECSF10, and NKCL, belongs to the C-type lectin family of transmembrane immune regulatory glycoproteins. Dectin-2alpha, CLEC4A/DCIR, CLEC4B/DCAR, CLEC4C/DLEC, CLEC4D/MCL, and CLEC4E/mincle constitute a subgroup of these molecules that exhibit approximately 40% aa sequence identity in their extracellular domains (ECD) and have a conserved cysteine spacing in their carbohydrate recognition domains (CRD). 1,2 Mature mouse dectin-2alpha consists of a short cytoplasmic tail, a transmembrane segment, and an ECD with a stalk region and one CRD. 3,4 Alternate splicing leads to partial deletion of the transmembrane segment and stalk (beta isoform) or a portion of the CRD (gamma isoform). 4 The full length dectin-2alpha isoform is a 27 kD molecule that is primarily expressed on the surface of tissue macrophages and their precursors. 3-6 The CRD of dectin-2alpha contains an EPN motif which is characteristic of calcium-dependent mannose-binding lectins. Dectin-2alpha selectively interacts with high mannose structures in the Man9GlcNAc2 configuration. 7 It mediates the recognition of a variety of microorganisms, particularly the filamentous forms of yeast and fungii. 7,8 The short cytoplasmic tail does not contain signaling motifs but mediates association with the ITAM-containing Fc receptor gamma subunit in macrophages. 8 Ligation of dectin-2alpha induces tyrosine phosphorylation of the gamma subunit, activation of NFkappaB, and enhanced release of TNF-alpha and IL-1ra. 8 Macrophage dectin-2alpha is upregulated in vivo by inflammatory stimuli and UV-B irradiation. 6 It mediates the breaking of UV-induced tolerance by interacting with CD4+ CD25+ regulatory T cells which then induce dendritic cells to release IL-4, IL-10, and TGF-beta. 9 Within the ECD, mouse dectin-2alpha shares 71% aa sequence identity with human and bovine dectin-2. Source: Human CD33 signal peptide (Met 1-Ala 16), HHHHHHHA, Mouse Dectin-2alpha (Phe43-Leu209). A DNA sequence encoding the N-terminal polyhistidine tagged extracellular domain of mouse dectin-2alpha (Phe 43-Leu 209, Accession # AAF67177) (Ariizumi, K., et al., 2000 J. Biol. Chem., 275: 11957-63) was fused to the signal peptide of human CD33 at the N-terminus. The protein was expressed in a mouse myeloma cell line, NS0. Molecular Mass: The recombinant mature polyhistidine tagged mouse dectin-2alpha, generated by the removal of the signal peptide region, is a monomeric protein. Based on N-terminal sequencing, the mature recombinant mouse dectin-2alpha begins with the histidine tag and has a calculated molecular mass of 20.6 kD. As a result of glycosylation, the recombinant protein migrates as an approximately 24 kD protein in SDS- PAGE under reducing conditions. Endotoxin Level: < 1.0 EU per 1 µg of the protein as determined by the LAL method. Activity: Measured by its ability to bind biotinylated alpha-D-Mannose-PAA in a functional ELISA. Reconstitution: It is recommended that sterile PBS be added to the vial to prepare a working stock solution of no less than 100ug/ml. The carrier-free protein should be used immediately upon reconstitution to avoid losses in activity due to non-specific binding to the inside surface of the vial. For long term storage as a dilute solution, a carrier protein (e.g. 0.1% HSA or BSA) should be added to the vial. Storage and Stability: Lyophilized samples are stable for up to twelve months at -20°C. Upon reconstitution, this protein, in the presence of a carrier protein, can be stored under sterile conditions at 2-8°C for one month or at -20°C in a manual defrost freezer for three months without detectable loss of activity. Avoid repeated freeze-thaw cycles.