Detection of Mouse Siglec‑E by Western Blot.
Western blot shows lysates of mouse spleen tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Siglec‑E Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5806) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Siglec‑E at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Siglec‑E in Mouse Spleen.
Siglec‑E was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse Siglec‑E Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5806) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
are sialic acid specific I‑type lectins that are characterized by an
extracellular domain (ECD) with an N‑terminal Ig‑like V‑set domain
followed by varying numbers of Ig‑like C2‑set domains (1, 2). Mouse
Siglec‑E, also known as Myeloid Inhibitory Siglec (MIS), is an
80 ‑ 85 kDa member of the CD33‑related subfamily of Siglecs. It consists
of a 335 amino acid (aa) ECD with one Ig‑like V‑set domain and two
Ig‑like C2‑set domains, a 21 aa transmembrane segment, and a 93 aa
cytoplasmic domain that contains two immunoreceptor tyrosine‑based
inhibitory motifs (ITIM) (3, 4). Rodent and primate Siglec gene families
have significantly diverged, and Siglec‑9 is the most likely human
ortholog of mouse Siglec‑E (1). Within the ECD, mouse Siglec‑E shares
56% and 80% aa sequence identity with human Siglec‑9 and rat Siglec‑E,
respectively. Siglec‑E is expressed as a heavily N‑glycosylated
disulfide‑linked homodimer and shows binding preference for disialic
acids in the alpha 2‑8 linkage (3, 5). It is expressed on the surface of
several hematopoietic cell types including neutrophils, NK cells,
monocytes, peritoneal macrophages and B1 cells, and splenic myeloid
dendritic cells and marginal zone B cells (5). Tyrosine phosphorylation
of the cytoplasmic ITIMs mediates the association of Siglec‑E with the
phosphatases SHP‑1 and SHP‑2 (3, 4). Siglec‑E is up‑regulated and
additionally phosphorylated following cellular stimulation by a variety
of TLR agonists (6). Siglec‑E signaling negatively regulates the
LPS‑induced production of TNF‑ alpha and IL‑6 by macrophages (4, 6). Its
up‑regulation in macrophages parallels the development of endotoxin
tolerance (6). Siglec‑E recognition of sialylated determinants on
virulent T. cruzi contributes to the suppression of dendritic cell IL‑12 p40 production (7).
Varki, A. and T. Angata (2006) Glycobiology 16:1R.
Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
Yu, Z. et al. (2001) Biochem. J. 353:483.
Ulyanova, T. et al. (2001) J. Biol. Chem. 276:14451.
Zhang, J.Q. et al. (2004) Eur. J. Immunol. 34:1175.
Boyd, C.R. et al. (2009) J. Immunol. 183:7703.
Erdmann, H. et al. (2009) Cell. Microbiol. 11:1600.
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