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Mouse Siglec-1/CD169 Antibody

Catalog #: AF5610 Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF5610
AF5610-SP
Detection of Mouse Siglec‑1/CD169 by Western Blot.
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Mouse Siglec-1/CD169 Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.

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.

Scientific Data

Western Blot Detection of Mouse Siglec-1/CD169 antibody by Western Blot. View Larger

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.

Neutralization Cell Adhesion Mediated by Siglec‑1/CD169 and Neutral-ization by Mouse Siglec‑1/CD169 Antibody. View Larger

Cell Adhesion Mediated by Siglec‑1/CD169 and Neutral-ization 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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
CD169; FLJ00051; sialic acid binding Ig-like lectin 1, sialoadhesin; sialoadhesin; Siglec1; Siglec-1

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