Siglec-2/CD22 Antibody [Unconjugated]

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.

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.

• Reactivity: Mu
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Siglec-2/CD22 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Siglec‑2/CD22
  Ser22-Arg702
  Accession # P35329
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Cd22
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 0.1-0.5 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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC 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-2/CD22 Antibody [Unconjugated]

• 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

Background

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).

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.

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-2/CD22 Antibody (AF2296)(1)

Publication using AF2296

• Horikawa K, Martin SW, Pogue SL, Silver K, Peng K, Takatsu K, Goodnow CC Enhancement and suppression of signaling by the conserved tail of IgG memory-type B cell antigen receptors. J. Exp. Med., 2007-04-09;204(4):759-69. 2007-04-09 [PMID: 17420266] (Western Blot, Mouse)

FAQs for Siglec-2/CD22 Antibody (AF2296). (Showing 1 - 1 of 1 FAQ).

1. I am interested in buying a CD22 antibody for fluorescence microscopy and FACS I red on your web page that you also offer to label your antibodies, if required, is that correct?
   • We do have the ability to perform custom conjugations on certain antibodies, including some to the CD22 protein. If you send me the conjugate you are interested in I can provide you with a quote.

Bioinformatics

• Gene Symbol: Cd22
• Uniprot:
  • P35329()