Human Siglec-1/CD169 Antibody

Catalog #: AF5197
3 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF5197
AF5197-SP

Detection of Human Siglec‑1/CD169 by Western Blot.

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Citations (3)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Siglec-1/CD169 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Siglec‑1/CD169 in direct ELISAs and Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Siglec‑1/CD169
Ser20-Gln1641
Accession # Q9BZZ2

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.5 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 1.5-7.5 µg/mL in the presence of 5 µg/mL Recombinant Human 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

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Detection of Human Siglec‑1/CD169 by Western Blot. 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.

Neutralization

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Cell Adhesion Mediated by Siglec‑1/CD169 and Neutralization by Human Siglec‑1/CD169 Antibody. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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). 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).

References

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

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Related Research Areas

Citations for Human Siglec-1/CD169 Antibody

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.

3 Citations: Showing 1 - 3
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Gene Expression Profiling of Lymph Node Sub-Capsular Sinus Macrophages in Cancer
Authors: Danilo Pellin, Natalie Claudio, Zihan Guo, Tahereh Ziglari, Ferdinando Pucci
Frontiers in Immunology
Distinct patterns and prognostic values of tumor-infiltrating macrophages in hepatocellular carcinoma and gastric cancer
Authors: JQ Li, XJ Yu, YC Wang, LY Huang, CQ Liu, L Zheng, YJ Fang, J Xu
J Transl Med, 2017-02-15;15(1):37.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P
Siglec-1 initiates formation of the virus-containing compartment and enhances macrophage-to-T cell transmission of HIV-1
Authors: JE Hammonds, N Beeman, L Ding, S Takushi, AC Francis, JJ Wang, GB Melikyan, P Spearman
PLoS Pathog, 2017-01-27;13(1):e1006181.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry