Human Siglec-5/Siglec-14 Antibody

(3 citations)   
  • Species Reactivity
    Human
  • 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
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Siglec-5
    Glu17-Thr434
    Accession # O15389
  • 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
    0.1 µg/mL
    Recombinant Human Siglec‑5 Fc Chimera (Catalog # 1072-SL)
  • Flow Cytometry
    2.5 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    Immersion fixed paraffin-embedded sections of human lymph node
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Neutralization
    Measured by its ability to neutralize Siglec‑5-mediated adhesion of human red blood cells. Kelm, S. et al. (1994) Current Biology 4:965. The Neutralization Dose (ND50) is typically 0.5-2.0 µg/mL in the presence of 5 µg/mL Recombinant Human Siglec‑5 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
Cell Adhesion Mediated by Siglec‑5 and Neutralization by Human Siglec‑5/14 Antibody. 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.
Detection of Siglec‑5/14 in Human Monocytes by Flow Cytometry. 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).
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-5/Siglec-14

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.

  • References:
    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.
  • Long Name:
    Sialic Acid Binding Ig-like Lectin 5
  • Entrez Gene IDs:
    8778 (Human)
  • Alternate Names:
    Siglec-5/Siglec-14
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  1. Soluble Siglec-5 associates to PSGL-1 and displays anti-inflammatory activity
    Sci Rep, 2016;6(0):37953.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  2. Siglec-9 enhances IL-10 production in macrophages via tyrosine-based motifs.
    Authors: Ando M, Tu W, Nishijima K, Iijima S
    Biochem. Biophys. Res. Commun., 2008;369(3):878-83.
    Species: Mouse
    Sample Type: Whole Cells
  3. IL-1F5 mediates anti-inflammatory activity in the brain through induction of IL-4 following interaction with SIGIRR/TIR8.
    Authors: Costelloe C, Watson M, Murphy A, McQuillan K, Loscher C, Armstrong ME, Garlanda C, Mantovani A, O'Neill LA, Mills KH, Lynch MA
    J. Neurochem., 2008;105(5):1960-9.
    Species: Rat
    Sample Type: Whole Cells
    Application: ICC
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