Mouse Siglec-2/CD22 Antibody

(1 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Siglec‑2/CD22 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross‑reactivity with recombinant human (rh) Siglec-2 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Siglec‑2/CD22
    Ser22-Arg702
    Accession # P35329
  • 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 Mouse Siglec‑2/CD22 Fc Chimera (Catalog # 2296-SL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize Siglec‑2/CD22-mediated adhesion of human red blood cells. Kelm, S. et al. (1994) Current Biology 4:965. The Neutralization Dose (ND50) is typically 0.1-0.5 µg/mL in the presence of 0.3 µg/mL Recombinant Mouse Siglec‑2/CD22 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‑2/CD22 and Neutral­ization by Mouse Siglec‑2/
CD22 Antibody.
Recombinant Mouse Siglec‑2/
CD22 Fc Chimera (Catalog # 2296-SL), immobilized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Siglec‑2/
CD22 Fc Chimera (0.3 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti‑Mouse Siglec‑2/CD22 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2296). The ND50 is typically 0.1-0.5 µg/mL.
Immunocytochemistry
Siglec‑2/CD22 in Mouse Splenocytes. Siglec‑2/CD22 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse Siglec‑2/CD22 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2296) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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-2/CD22

Siglecs (sialic acid binding Ig-like lectins) are I-type (Ig-type) lectins belonging to the Ig superfamily. They are characterized by an N‑terminal Ig-like V-type domain which mediates sialic acid binding, followed by varying numbers of Ig-like C2-type domains (1, 2). Eleven human Siglecs have been cloned and characterized. Among these are sialoadhesin/CD169/Siglec-1, CD22/Siglec-2 and CD33/Siglec-3. To date, no Siglec has been shown to recognize any cell surface ligand other than sialic acid, suggesting that interactions with glycans containing this carbohydrate are important in mediating the biological functions of Siglecs. The cDNA of mouse Siglec-2 (also known as B-cell antigen CD22), encodes an 862 amino acid (aa) protein that contains a 21 aa signal peptide, a 681 aa extracellular region, a 19 aa transmembrane region and a 141 aa cytoplasmic tail (3, 4). The extracellular region contains one N-terminal V-type Ig-like domain followed by six Ig-like C2-type domains. The cytoplasmic domain has 3 immunoreceptor tyrosine-based inhibition motifs (ITIMs). Two splice forms exist, both showing deletions in the V-type Ig domain of 30 aa and 60 aa each. There are also two alleles in mouse that account for a difference of 10 aa in the extracellular region. The extracellular region of mouse Siglec-2 is 60% aa identity to human extracellular Siglec-2. Expression of mouse Siglec-2/CD22 generates a 140 kDa integral membrane glycoprotein that is limited to the B cell compartment of lymphoid tissues. Its expression is upregulated by LPS activtion (5, 6). Siglec-2/CD22 is an adhesion molecule that preferentially binds alpha 2,6- linked sialic acid on the same (cis) or adjacent (trans) cells. Interaction of CD22 with trans ligands on opposing cells was found to be favored over the binding of ligands in cis (7).

  • References:
    1. Nitschke, L. et al. (2001) Scand. J. Immunol. 53:227.
    2. Crocker, P.R. and A. Varki (2001) Immunology 103:137.
    3. Law, C-L. et al. (1993) J. Immunol. 151:175.
    4. Wienands, Y.J. et al. (1999) J. Biol. Chem. 274:18769.
    5. Wilson, G.L et al. (1991) J. Exp. Med. 173:137.
    6. Torres, P.M. (1992) J. Immunol. 194:2641.
    7. Collins, B.E. et al. (2004) Proc. Natl. Acad. Sci. 101:6104.
  • Long Name:
    Sialic Acid Binding Ig-like Lectin 2
  • Entrez Gene IDs:
    933 (Human); 12483 (Mouse); 102130968 (Cynomolgus Monkey)
  • Alternate Names:
    B-cell receptor CD22; BL-CAM; B-lymphocyte cell adhesion molecule; CD22 antigenMGC130020; CD22 molecule; CD22; sialic acid binding Ig-like lectin 2; Sialic acid-binding Ig-like lectin 2; Siglec2; Siglec-2; SIGLEC2FLJ22814; T-cell surface antigen Leu-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.

1 Citations: Showing 1 - 1

  1. Enhancement and suppression of signaling by the conserved tail of IgG memory-type B cell antigen receptors.
    Authors: Horikawa K, Martin SW, Pogue SL, Silver K, Peng K, Takatsu K, Goodnow CC
    J. Exp. Med., 2007;204(4):759-69.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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