Detection of Siglec‑11 in HEK293 Human Cell Line Transfected with Human Siglec-11 and eGFP by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with either (A) human Siglec-11 or (B) human Siglec-16 and eGFP was stained with Mouse Anti-Human Siglec‑11 Monoclonal Antibody (Catalog # MAB3258) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and. Quadrant markers were set based on control antibody staining (Catalog # MAB003). View our protocol for Staining Membrane-associated Proteins.
Siglec‑11 in Human Lymph Node.
Siglec‑11 was detected in immersion fixed paraffin-embedded sections of human lymph node using Mouse Anti-Human Siglec‑11 Monoclonal Antibody (Catalog # MAB3258) at 5 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Siglecs (sialic acid binding Ig-like lectins) are I-type lectins that belong to the immunoglobulin superfamily. They are characterized by an N‑terminal Ig-like V-set domain which mediates sialic acid binding, followed by a varying numbers of Ig-like C2-set domains. Siglecs‑3 and 5‑13 constitute the CD33/Siglec-3 related group, which are defined by their sequence homology and differential expression in the hematopoietic system (1‑3). Mature human Siglec-11 consists of a 534 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 114 aa cytoplasmic domain. The ECD contains one Ig-like V-set domain, and three Ig-like C2-set domains. The cytoplasmic domain contains two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) (4). A splice variant of Siglec-11 has a deletion of nearly 100 aa in the extracellular juxtamembrane region. Among siglecs, the ECD of Siglec-11 is most closely related to that of Siglec-10 (82% aa sequence identity). The cytoplasmic domains of these proteins are only 20% identical. Siglec-11 is closely related to the pseudogenes Siglec-14 and Siglec-16 (4, 5). Human Siglec-11 shares 90%‑96% aa sequence identity with Siglec-11 from great apes. Rodent orthologs of Siglec-11 have not been identified. In human, Siglec-11 is expressed in tissue macrophages, brain microglia, and inflammatory site monocytes (4). Strong microglial expression is specific to humans, as it is less prominent or absent in chimpanzees and orangutans (5). Siglec-11 forms 180 kDa disulfide-linked dimers. It shows a strong binding preference for sialic acid in alpha 2-8 linkage which is unusual for siglecs (4). A conserved arginine in the Ig-like V-set domain only partially contributes to Siglec-11 ligand recognition, in contrast to its being required in other siglecs (4). Tyrosine phosphorylation of the cytoplasmic region of Siglec-11 promotes association with the phosphatases SHP-1 and SHP-2 (4).
Varki, A. and T. Angata (2006) Glycobiology 16:1R.
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