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Human Siglec-5/Siglec-14 Antibody

Catalog #: AF1072
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1072
AF1072-SP
Cell Adhesion Mediated by Siglec‑5 and Neutralization by Human Siglec‑5/14 Antibody.
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Citations (6)
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Human Siglec-5/Siglec-14 Antibody Summary

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 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
0.1 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 tonsil tissue
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.

Scientific Data

Neutralization Cell Adhesion Mediated by Siglec‑5 and Neutralization by Human Siglec‑5/14 Antibody. View Larger

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.

Flow Cytometry Detection of Siglec-5/14 antibody in Human Monocytes antibody by Flow Cytometry. View Larger

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).

Immunohistochemistry View Larger

Siglec‑5/Siglec‑14 in Human Tonsil. 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.

Reconstitution Calculator

Reconstitution Calculator

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

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Citations for Human Siglec-5/Siglec-14 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.

6 Citations: Showing 1 - 6
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  1. Novel Virulence Role of Pneumococcal NanA in Host Inflammation and Cell Death Through the Activation of Inflammasome and the Caspase Pathway
    Authors: Yu-Wen Tseng, Chun-Chi Chang, Yung-Chi Chang
    Frontiers in Cellular and Infection Microbiology
  2. Decidual glycodelin-A polarizes human monocytes towards a decidual macrophage-like phenotype via siglec-7
    Authors: M Vijayan, CL Lee, VHH Wong, X Wang, K Bai, J Wu, H Koistinen, M Seppälä, KF Lee, WSB Yeung, EHY Ng, PCN Chiu
    J. Cell. Sci., 2020-07-23;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. Siglec expression on the surface of human, bull and ram sperm
    Authors: 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
    Reproduction, 2018-04-01;155(4):361-371.
    Species: Bovine, Human, Ovine
    Sample Types: Tissue Homogenates, Whole Cells
    Applications: IHC, Western Blot
  4. Soluble Siglec-5 associates to PSGL-1 and displays anti-inflammatory activity
    Sci Rep, 2016-11-28;6(0):37953.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  5. 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., 2008-03-04;369(3):878-83.
    Species: Mouse
    Sample Types: Whole Cells
  6. 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., 2008-02-18;105(5):1960-9.
    Species: Rat
    Sample Types: Whole Cells
    Applications: ICC, Neutralization

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