Recombinant Human Siglec-2/CD22 Fc Alexa Fluor® 647 Protein

Fc Chimera
Catalog # Availability Size / Price Qty
AFR1968-020
Flow cytometry analysis for Recombinant Human Siglec‑2/CD22 Fc Chimera Alexa Fluor® 647 staining on anti-human Siglec‑2/CD22 Monoclonal Antibody conjugated beads.
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Recombinant Human Siglec-2/CD22 Fc Alexa Fluor® 647 Protein Summary

Learn more about FluorokinesTM Fluorescent-Labeled Proteins

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by flow cytometry for its ability to bind anti-human Siglec‑2/CD22 Monoclonal Antibody conjugated beads.The concentration of Recombinant Human Siglec‑2/CD22 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1968) that produces 50% of the binding response is 2.50‑25.0 ng/mL
Source
Mouse myeloma cell line, NS0-derived human Siglec-2/CD22 protein
Human Siglec-2
(Asp20-Arg687)
Accession # CAA42006.1
DIEGRMD Human IgG1
(Pro100-Lys330)
N-terminus C-terminus
Accession #
N-terminal Sequence
Analysis
Asp20
Structure / Form
Disulfide-linked homodimer
Labeled with Alexa Fluor® 647
Excitation Wavelength: 650 nm
Emission Wavelength: 668 nm
Predicted Molecular Mass
101.9 kDa (monomer)
SDS-PAGE
120-132 kDa, under reducing conditions.

Product Datasheets

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AFR1968

AFR1968

Formulation Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Scientific Data

Flow Cytometry View Larger

Streptavidin coated beads conjugated to biotinylated anti-human Siglec‑2/CD22 Monoclonal Antibody were stained with the indicated concentrations of Recombinant Human Siglec‑2/CD22 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1968).

SDS-PAGE View Larger

2 μg/lane of Recombinant Human Siglec‑2/CD22 Fc Chimera Alexa Fluor® 647 Protein (Catalog # AFR1968) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 120-132 kDa and 240-260 kDa, respectively.

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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. They are sialoadhesin/CD169/Siglec-1, CD22/Siglec-2, CD33/Siglec-3, Myelin-Associated Glycoprotein (MAG/Siglec-4a) and the identified Siglecs 5 to 11 (1 ‑ 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. Human Siglec-2, also known as B-cell antigen CD22 or B-lymphocyte cell adhesion molecule (BL-CAM), is a B-cell restricted glycoprotein that is expressed in the cytoplasm of progenitor B and pre-B cells and on the surface of mature B cells. Two distinct human Siglec-2/CD22 cDNAs that arise from differential RNA processing of the same gene have been isolated. The predominant Siglec-2/CD22 beta encodes an 847 amino acid (aa) polypeptide with a hydrophobic signal peptide, an N-terminal Ig-like V-type domain, six Ig-like C2-type domains, a transmembrane region and a cytoplasmic tail with 4 immunoreceptor tyrosine-based inhibition motifs (ITIMs) (4). The variant Siglec-2/CD22 alpha encodes a 647 aa polypeptide missing two Ig-like C2-type domains and has a truncated (23 aa) cytoplasmic tail (5). 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 (9). Besides its role as an adhesion molecule, Siglec-2/CD22 is a coreceptor that physically interacts with B-cell receptor (BCR) and is rapidly phosphorylated upon BCR ligation. It negatively regulates BCR signals by recruiting tyrosine phosphatase SHP-1 to its ITIMs. Phosphorylated Siglec-2/CD22 can also interact with other intracellular effector proteins such as Syk, PLC gamma, PI3 kinase and Grb-2, suggesting it may play a role in positive signaling (2, 7, 8).

References
  1. Crocker, P.R. and A. Varki (2001) Trends Immunol. 22:337.
  2. Crocker, P.R. and A. Varki (2001) Immunology 103:137.
  3. Angata, T. et al. (2002) J. Biol. Chem. 277:24466.
  4. Wilson, G.L et al. (1991) J. Exp. Med. 173:137.
  5. Stamenkovic, I. and B. Seed (1990) Nature 345:74.
  6. Kelm, S. et al. (1994) Current Bio. 4:965.
  7. Ravetch, J.V. and L.L. Lanier (2000) Science 290:84.
  8. Wienands, Y.J. et al. (1999) J. Biol. Chem. 274:18769.
  9. 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); 308501 (Rat); 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

Product Specific Notices

This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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