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Recombinant Human Siglec-3/CD33 Protein, Atto 488 Conjugate

Catalog #: ATJ1137 Datasheet / COA / SDS
Fc Chimera
Catalog # Availability Size / Price Qty
ATJ1137-050
Detection of Human Siglec-3/CD33 Antibody with Recombinant Human Siglec-3/CD33 Protein, Atto 488 Conjugate by Flow Cytometry.
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Recombinant Human Siglec-3/CD33 Protein, Atto 488 Conjugate Summary

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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 fluorescent beads.

Source
Mouse myeloma cell line, NS0-derived human Siglec-3/CD33 protein
Human Siglec-3/CD33
(Asp18-His259)
(Val257Leu)
Accession # AAA51948.1		DIEGRMD Human IgG1
(Pro100-Lys330)
N-terminusC-terminus
Accession #
AAA51948.1
N-terminal Sequence
Analysis
Asp18
Structure / Form
Disulfide-linked homodimer, labeled with Atto 488 via amines
Excitation Wavelength: 501 nm
Emission Wavelenght: 523 nm
Predicted Molecular Mass
53.4 kDa (monomer)
SDS-PAGE
67-80 kDa, under reducing conditions

Product Datasheets

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ATJ1137

Product Datasheet
COA
SDS

ATJ1137

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

Fluorescent beads conjugated to anti-Human Siglec 3/CD33 Monoclonal Antibody (MAB1137) were stained with (A) Recombinant Human Siglec 3/CD33 Protein, Atto 488 Conjugate (Catalog # ATJ1137) or (B) unstained.

SDS-PAGE View Larger

2 μg/lane of Recombinant Human Siglec-3/CD33 Protein, Atto 488 Conjugate (Catalog # ATJ1137) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 67-80 kDa and 134-160 kDa, respectively.

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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

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