Siglec-5/Siglec-14 Antibody [Unconjugated]

Recombinant Human Siglec‑5 Fc Chimera (Catalog # 1072-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‑5 Fc Chimera (5 µg/mL) is neutralized (green line) by increasing concentrations of Human Siglec‑5/14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1072). The ND50 is typically 0.5-2.0 µg/mL.

Human whole blood monocytes were stained with Human Siglec‑5/14 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1072, filled histogram) or control antibody (Catalog # AB‑108‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

Siglec‑5/Siglec‑14 was detected in immersion fixed paraffin-embedded sections of human tonsil tissue using Goat Anti-Human Siglec‑5/Siglec‑14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1072) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes in germinal centers. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Flow, IHC, B/N, CyTOF-ready
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Siglec-5/Siglec-14 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Siglec-5
  Glu17-Thr434
  Accession # O15389
• Specificity: Detects human Siglec-5/14 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human (rh) Siglec-7 and rhSiglec-9 is observed and less than 1% cross-reactivity with rhSiglec-2 and rhSiglec-3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SIGLEC5
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 0.5-2.0 ug/mL
  • Western Blot 0.1 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-5/Siglec-14 Antibody [Unconjugated]

• CD170;CD33L2;OBBP2;OB-BP2;Sialic acid-binding Ig-like lectin 5;SIGLEC5;SIGLEC-5
• Siglec-5/Siglec-14

Background

Siglecs (1) (sialic acid binding Ig-like lectins) are I-type (Ig-type) lectins (2) belonging to the Ig superfamily. They are characterized by an N-terminal Ig-like V-type domain which mediates sialic acid binding (3), followed by varying numbers of Ig-like C2-type domains (1, 4). Eleven human Siglecs have been cloned and characterized (1, 4). They are sialoadhesin/CD169/Siglec-1, CD22/Siglec-2, CD33/Siglec-3, Myelin-Associated Glycoprotein (MAG/Siglec-4a) and the Siglec-5 to 11 (4, 5,  6). To date, no Siglec has been shown to recognized any cell surface ligand other than sialic acids, suggesting that interactions with glycans containing this carbohydrate are important in mediating the biological functions of Siglecs. Siglec-5 to 11 share a high degree of sequence similarity with CD33/Siglec-3 both in their extracellular and intracellular regions. They are collectively referred to as CD33-related Siglecs. One remarkable feature of the CD33-related Siglecs is their differential expression pattern within the hematopoietic system (4, 5). This fact, together with the presence of two conserved immunoreceptor tyrosine-based inhibition motifs (ITIMs) in their cytoplasma tails, suggests that CD33-related Siglecs are involved in the regulation of cellular activation within the immune system.

1. Crocker, P.R. et al. (1998) Glycobiology 8:v.
2. Powell, L.D. et al. (1995) J. Biol. Chem. 270:14243.
3. May, A.R. et al. (1998) Mol. Cell 1998. 1:719.
4. Crocker, P.R. and A. Varki (2001) Trends Immunol. 22:337.
5. Crocker, P.R. et al. (2001) Immunology 103:137.
6. Angata, T. et al. (2002) J. Biol Chem. 277:24466.

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-5/Siglec-14 Antibody (AF1072)(7)

Publications using AF1072

• Yu-Wen Tseng, Chun-Chi Chang, Yung-Chi Chang Novel Virulence Role of Pneumococcal NanA in Host Inflammation and Cell Death Through the Activation of Inflammasome and the Caspase Pathway Frontiers in Cellular and Infection Microbiology 2021-03-11 [PMID: 33777832] (Western Blot, Human)
• Yu-Wen Tseng, Chun-Chi Chang, Yung-Chi Chang Novel Virulence Role of Pneumococcal NanA in Host Inflammation and Cell Death Through the Activation of Inflammasome and the Caspase Pathway Frontiers in Cellular and Infection Microbiology 3/11/2021 [PMID: 33777832]
• M Vijayan, CL Lee, VHH Wong, X Wang, K Bai, J Wu, H Koistinen, M Seppälä, KF Lee, WSB Yeung, EHY Ng, PCN Chiu Decidual glycodelin-A polarizes human monocytes towards a decidual macrophage-like phenotype via siglec-7 J. Cell. Sci., 2020-07-23;0(0):. 2020-07-23 [PMID: 32513821] (Neutralization, Human)
• K Alkhodair, H Almhanna, J McGetrick, S Gedair, ME Gallagher, B Fernandez-, T Tharmaling, PB Larsen, E Fitzpatric, P Lonergan, ACO Evans, SD Carrington, CJ Reid Siglec expression on the surface of human, bull and ram sperm Reproduction, 2018-04-01;155(4):361-371. 2018-04-01 [PMID: 29581386] (IHC, Western Blot, Bovine, Human, Ovine)
• Soluble Siglec-5 associates to PSGL-1 and displays anti-inflammatory activity Sci Rep, 2016-11-28;6(0):37953. 2016-11-28 [PMID: 27892504] (ICC, Human)
• Ando M, Tu W, Nishijima K, Iijima S Siglec-9 enhances IL-10 production in macrophages via tyrosine-based motifs. Biochem. Biophys. Res. Commun., 2008-03-04;369(3):878-83. 2008-03-04 [PMID: 18325328] (Mouse)
• Costelloe C, Watson M, Murphy A, McQuillan K, Loscher C, Armstrong ME, Garlanda C, Mantovani A, O'Neill LA, Mills KH, Lynch MA IL-1F5 mediates anti-inflammatory activity in the brain through induction of IL-4 following interaction with SIGIRR/TIR8. J. Neurochem., 2008-02-18;105(5):1960-9. 2008-02-18 [PMID: 18284608] (ICC, Neutralization, Rat)

Bioinformatics

• Gene Symbol: SIGLEC5
• Uniprot:
  • O15389(Human)