Recombinant Human Siglec-3/CD33 Fc Alexa Fluor® 647 Protein

Catalog # Availability Size / Price Qty
AFR1137-020
Flow cytometry analysis for Recombinant Human Siglec‑3/CD33 Fc Chimera Alexa Fluor® 647 staining on anti-human Siglec‑3/CD33 Monoclonal Antibody conjugated beads.
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Recombinant Human Siglec-3/CD33 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‑3/CD33 Monoclonal Antibody conjugated beads.The concentration of Recombinant Human Siglec‑3/CD33 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1137) that produces 50% of the binding response is 0.50‑20.0 ng/mL.
Source
Mouse myeloma cell line, NS0-derived human Siglec-3/CD33 protein
Human Siglec-3
(Asp18-His259)(Val257Leu)
Accession # AAA51948
DIEGRMD Human IgG1
(Pro100-Lys330)
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Asp18
Structure / Form
Disulfide-linked homodimer
Labeled with Alexa Fluor® 647 via amines
Excitation Wavelength: 650 nm
Emission Wavelength: 668 nm
Predicted Molecular Mass
53.4 kDa (monomer)
SDS-PAGE
67-85 kDa, under reducing conditions

Product Datasheets

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AFR1137

AFR1137

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‑3/CD33 Monoclonal Antibody were stained with the indicated concentrations of Recombinant Human Siglec‑3/CD33 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1137).

SDS-PAGE View Larger

2 μg/lane of Recombinant Human Siglec‑3/CD33 Fc Chimera Alexa Fluor® 647 Protein (Catalog # AFR1137) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 67-85 kDa.

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Background: Siglec-3/CD33

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 Siglecs 5 to 11 (1 - 3). 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. Siglecs 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 (1, 2). 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. Human Siglec-3 is alternatively known as myeloid cell surface antigen CD33 and GP67. Human Siglec-3 cDNA encodes a 364 amino acid (aa) polypeptide with a hydrophobic signal peptide, an N-terminal Ig-like V-type domain, one Ig-like C2-type domains, a transmembrane region and a cytoplasmic tail (1, 4). Siglec-3 expression is restricted to cells of myelomonocytic lineage (2). It binds sialic acid preferring alpha 2,3- linkage over alpha 2,6- linkage (5). Studies indicated that Siglec-3 recruits SHP-1 and SHP-2 to its ITIMs (6, 7). When co-crosslinking with Fc gamma R1, Siglec-3 inhibits tyrosine phosphorylation and calcium mobilization, suggesting Siglec-3 can mediate inhibitory signals (7).

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. Simmons, D. and B. Seed (1988) J. Immunol. 141:2797.
  5. Freeman, S.D. et al. (1995) Blood 85:2002.
  6. Taylor, V.C. et al. (1999) J. Biol. Chem. 274:11505.
  7. Ulyanova, T. et al. (1999) Eur. J. Immunol. 29:3440.
Long Name
Sialic Acid Binding Ig-like Lectin 3
Entrez Gene IDs
945 (Human); 12489 (Mouse); 102117580 (Cynomolgus Monkey)
Alternate Names
CD33 antigen (gp67); CD33 antigen; CD33 molecule; CD33; FLJ00391; gp67; myeloid cell surface antigen CD33; p67; sialic acid binding Ig-like lectin 3; Sialic acid-binding Ig-like lectin 3; Siglec3; Siglec-3; SIGLEC3gp67

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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