Siglec-1/CD169 Antibody [Unconjugated]

Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line and human lymph node. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Siglec‑1/CD169 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5197) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Siglec‑1/CD169 at approximately 180-200 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Recombinant Human Siglec‑1/CD169 Fc Chimera (Catalog # 5197-SL), immobilized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human Siglec‑1/CD169 Fc Chimera (5 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human Siglec‑1/CD169 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5197). The ND50 is typically 1.5-7.5 µg/mL.

• Reactivity: Hu
• Applications: WB, B/N
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Siglec-1/CD169 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Siglec‑1/CD169
  Ser20-Gln1641
  Accession # Q9BZZ2
• Specificity: Detects human Siglec‑1/CD169 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: SIGLEC1
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Neutralization 1.5-7.5 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Siglec-1/CD169 Antibody [Unconjugated]

• CD169
• FLJ00051
• sialic acid binding Ig-like lectin 1, sialoadhesin
• sialoadhesin
• Siglec1
• Siglec-1

Background

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). Human Siglec-1, also known as sialoadhesin and CD169, is a 175‑185 kDa glycoprotein. It contains a 1622 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 44 aa cytoplasmic domain (3). Within the ECD, human Siglec-1 shares approximately 70% aa sequence identity with mouse and rat Siglec-1. Alternate splicing generates a potentially soluble form of the ECD, and a second isoform with a substituted cytoplasmic domain. Siglec-1 expression is restricted to lymph node and splenic macrophages, plus some tissue macrophages (3). 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 (3‑5). Siglec-1 binds a number of sialylated molecules including the mannose receptor, MGL1, MUC1, PSGL-1, and different glycoforms of CD43 (6‑9). Its binding capacity can be masked by endogenous sialylated molecules (10, 11). The sialylated and sulfated N-linked carbohydrates that modify Siglec-1 itself are required for ligand binding (6, 7). Siglec-1 is expressed on dendritic cells following rhinovirus exposure, and these DC promote T cell anergy (12). It is also induced on circulating monocytes during systemic sclerosis and HIV-1 infection (13‑15). Siglec-1 can trap HIV-1 particles for trans infection of permissive cells (14).

1. Varki, A. and T. Angata (2006) Glycobiology 16:1R.
2. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
3. Hartnell, A. et al. (2001) Blood 97:288.
4. Nath, D. et al. (1995) J. Biol. Chem. 270:26184.
5. Crocker, P.R. et al. (1991) EMBO J. 10:1661.
6. Martinez-Pomares, L. et al. (1999) J. Biol. Chem. 274:35211.
7. Kumamoto, Y. et al. (2004) J. Biol. Chem. 279:49274.
8. Nath, D. et al. (1999) Immunology 98:213.
9. van den Berg, T.K. et al. (2001) J. Immunol. 166:3637.
10. Nakamura, K. et al. (2002) Glycobiology 12:209.
11. Barnes, Y.C. et al. (1999) Blood 93:1245.
12. Kirchberger, S. et al. (2005) J. Immunol. 175:1145.
13. York, M.R. et al. (2007) Arthritis Rheum. 56:1010.
14. Rempel, H. et al. (2008) PloS ONE 3:e1967.
15. van der Kuyl, A.C. et al. (2007) Plos ONE 2:e257.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Siglec-1/CD169 Antibody (AF5197)(4)

Publications using AF5197

• Danilo Pellin, Natalie Claudio, Zihan Guo, Tahereh Ziglari, Ferdinando Pucci Gene Expression Profiling of Lymph Node Sub-Capsular Sinus Macrophages in Cancer Frontiers in Immunology 6/8/2021 [PMID: 34168645]
• Danilo Pellin, Natalie Claudio, Zihan Guo, Tahereh Ziglari, Ferdinando Pucci Gene Expression Profiling of Lymph Node Sub-Capsular Sinus Macrophages in Cancer Frontiers in Immunology 2021-06-08 [PMID: 34168645] (Immunohistochemistry, Human)
• JQ Li, XJ Yu, YC Wang, LY Huang, CQ Liu, L Zheng, YJ Fang, J Xu Distinct patterns and prognostic values of tumor-infiltrating macrophages in hepatocellular carcinoma and gastric cancer J Transl Med, 2017-02-15;15(1):37. 2017-02-15 [PMID: 28202073] (IHC-P, Human)
• JE Hammonds, N Beeman, L Ding, S Takushi, AC Francis, JJ Wang, GB Melikyan, P Spearman Siglec-1 initiates formation of the virus-containing compartment and enhances macrophage-to-T cell transmission of HIV-1 PLoS Pathog, 2017-01-27;13(1):e1006181. 2017-01-27 [PMID: 28129379] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: SIGLEC1
• Uniprot:
  • Q9BZZ2(Human)