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

R&D Systems, part of Bio-Techne | Catalog # AFR1137

R&D Systems, part of Bio-Techne
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AFR1137-020

Key Product Details

Source

NS0

Accession #

Structure / Form

Disulfide-linked homodimer
Labeled with Alexa Fluor® 647 via amines
Excitation Wavelength: 650 nm
Emission Wavelength: 668 nm

Conjugate

Alexa Fluor 647 (Excitation= 650 nm, Emission= 668 nm)

Applications

Bioactivity

Product Specifications

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-terminus C-terminus

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.

N-terminal Sequence Analysis

Asp18

Predicted Molecular Mass

53.4 kDa (monomer)

SDS-PAGE

67-85 kDa, under reducing conditions

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.

Scientific Data Images for Recombinant Human Siglec-3/CD33 Fc Alexa Fluor® 647 Protein

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.

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).

Recombinant Human Siglec‑3/CD33 Fc Chimera Alexa Fluor® 647 Protein SDS-PAGE.

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.

Formulation, Preparation and Storage

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.

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 alpha2,3- linkage over alpha2,6- linkage (5). Studies indicated that Siglec-3 recruits SHP-1 and SHP-2 to its ITIMs (6, 7). When co-crosslinking with Fc gammaR1, 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

Alternate Names

CD33, gp67, Siglec3

Entrez Gene IDs

945 (Human); 12489 (Mouse); 102117580 (Cynomolgus Monkey)

Gene Symbol

CD33

UniProt

Additional Siglec-3/CD33 Products

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human Siglec-3/CD33 Fc Alexa Fluor® 647 Protein

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.

For research use only

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