Mouse Siglec-1/CD169 Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Siglec‑1/CD169 in direct ELISAs and Western blots. In direct ELISAs, approximately 30% cross-reactivity with recombinant human (rh) Siglec-1 is observed and less than 1% cross-reactivity with rhSiglec-2 and recombinant mouse Siglec-2 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Siglec‑1/CD169
    Thr20-Leu1639
    Accession # Q62230
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize Siglec‑1/CD169-mediated adhesion of human red blood cells. Kelm, S. et al. (1994) Current Biology 4:965. The Neutralization Dose (ND50) is typically 1-5 µg/mL in the presence of 5 µg/mL Recombinant Mouse Siglec‑1/CD169 Fc Chimera.
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‑1/CD169 by Western Blot. Western blot shows lysates of mouse lymph node tissue. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse Siglec‑1/CD169 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5610) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Siglec‑1/CD169 at approximately 180-190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Cell Adhesion Mediated by Siglec‑1/CD169 and Neutral­ization by Mouse Siglec‑1/CD169 Antibody.

Cell Adhesion Mediated by Siglec‑1/CD169 and Neutralization by Mouse Siglec‑1/CD169 Antibody. Recombinant Mouse Siglec‑1/CD169 Fc Chimera (Catalog # 5610-SL), immobilized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Siglec‑1/CD169 Fc Chimera (5 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Mouse Siglec‑1/CD169 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5610). The ND50 is typically 1-5 µg/mL.

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-1/CD169
Siglecs are sialic acid specific I-type lectins that belong to the immunoglobulin superfamily. Structurally, they are transmembrane proteins with an N-terminal Ig-like V‑set domain followed by varying numbers of Ig-like C2-set domains (1, 2). Mouse Siglec-1, also known as sialoadhesin and CD169, is a 175‑185 kDa glycoprotein that consists of a 1619 amino acid (aa) extracellular domain (ECD) with one Ig-like V‑set domain and 16 Ig-like C2-set domains, a 21 aa transmembrane segment, and a 35 aa cytoplasmic domain (3, 4). Within the ECD, mouse Siglec-1 shares 73% and 83% aa sequence identity with human and rat Siglec-1, respectively. Alternate splicing generates a soluble form of the ECD and a soluble isoform that is truncated following the first three Ig-like domains (3). Siglec-1 expression is restricted to lymph node and spleen macrophages and some tissue macrophages (4). The adhesive function of Siglec-1 is supported by the N-terminal Ig-like domain which shows a selectivity for alpha -2,3-linked sialic acid residues (4‑6). Siglec-1 binds a number of sialylated molecules including the mannose receptor, MGL1, MUC1, PSGL-1, and different glycoforms of CD43 (7‑10). Its binding capacity can be masked by endogenous sialylated molecules (11, 12). The sialylated and sulfated N-linked carbohydrates that modify Siglec-1 itself are required for ligand binding (7, 8). Siglec-1 is expressed on dendritic cells following rhinovirus exposure, and these DC promote T cell anergy (13). It is also induced on circulating monocytes during systemic sclerosis and HIV‑1 infection (14‑16). Siglec-1 can trap HIV‑1 particles for trans-infection of permissive cells (15).
  • References:
    1. Varki, A. and T. Angata (2006) Glycobiology 16:1R.
    2. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
    3. Crocker, P.R. et al. (1994) EMBO J. 13:4490.
    4. Hartnell, A. et al. (2001) Blood 97:288.
    5. Nath, D. et al. (1995) J. Biol. Chem. 270:26184.
    6. Crocker, P.R. et al. (1991) EMBO J. 10:1661.
    7. Martinez-Pomares, L. et al. (1999) J. Biol. Chem. 274:35211.
    8. Kumamoto, Y. et al. (2004) J. Biol. Chem. 279:49274.
    9. Nath, D. et al. (1999) Immunology 98:213.
    10. van den Berg, T.K. et al. (2001) J. Immunol. 166:3637.
    11. Nakamura, K. et al. (2002) Glycobiology 12:209.
    12. Barnes, Y.C. et al. (1999) Blood 93:1245.
    13. Kirchberger, S. et al. (2005) J. Immunol. 175:1145.
    14. York, M.R. et al. (2007) Arthritis Rheum. 56:1010.
    15. Rempel, H. et al. (2008) PloS ONE 3:e1967.
    16. van der Kuyl, A.C. et al. (2007) Plos ONE 2:e257.
  • Long Name:
    Sialic Acid Binding Ig-like Lectin 1
  • Entrez Gene IDs:
    6614 (Human); 20612 (Mouse); 311426 (Rat)
  • Alternate Names:
    C19orf75; CD169; Siglec1; Siglec-1; SIGLECL1 SIGLEC family like 1
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