Mouse Siglec-E Antibody

(4 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Siglec-E in direct ELISAs and Western blots. In direct ELISAs, less than 3% cross-reactivity with recombinant human (rh) Siglec-6, rhSiglec-7, and rhSiglec-9 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Siglec-E
    Gln20-Phe355
    Accession # Q6PJ50
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunohistochemistry
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
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: Siglec-E
Siglecs 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).
  • References:
    1. Varki, A. and T. Angata (2006) Glycobiology 16:1R.
    2. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
    3. Yu, Z. et al. (2001) Biochem. J. 353:483.
    4. Ulyanova, T. et al. (2001) J. Biol. Chem. 276:14451.
    5. Zhang, J.Q. et al. (2004) Eur. J. Immunol. 34:1175.
    6. Boyd, C.R. et al. (2009) J. Immunol. 183:7703.
    7. Erdmann, H. et al. (2009) Cell. Microbiol. 11:1600.
  • Long Name:
    Sialic Acid Binding Ig-like Lectin E
  • Entrez Gene IDs:
    83382 (Mouse)
  • Alternate Names:
    SiglecE; Siglec-E
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Siglec-7 restores ?-cell function and survival and reduces inflammation in pancreatic islets from patients with diabetes
    Authors: G Dharmadhik, K Stolz, M Hauke, NG Morgan, A Varki, E de Koning, S Kelm, K Maedler
    Sci Rep, 2017;7(0):45319.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  2. Leishmania donovani Utilize Sialic Acids for Binding and Phagocytosis in the Macrophages through Selective Utilization of Siglecs and Impair the Innate Immune Arm
    PLoS Negl Trop Dis, 2016;10(8):e0004904.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  3. Siglec receptors impact mammalian lifespan by modulating oxidative stress.
    Authors: Schwarz F, Pearce O, Wang X, Samraj A, Laubli H, Garcia J, Lin H, Fu X, Garcia-Bingman A, Secrest P, Romanoski C, Heyser C, Glass C, Hazen S, Varki N, Varki A, Gagneux P
    Elife, 2015;4(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  4. Group B Streptococcus engages an inhibitory Siglec through sialic acid mimicry to blunt innate immune and inflammatory responses in vivo.
    Authors: Chang Y, Olson J, Beasley F, Tung C, Zhang J, Crocker P, Varki A, Nizet V
    PLoS Pathog, 2014;10(1):e1003846.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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