Recombinant Human Siglec-3/CD33 Protein, Atto 647N Conjugate

Catalog #: ATM1137 Datasheet / COA / SDS

Fc Chimera

Catalog #	Availability	Size / Price	Qty
ATM1137-050

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Recombinant Human Siglec-3/CD33 Protein, Atto 647N 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) Accession # AAA51948.1	DIEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

AAA51948.1

N-terminal Sequence
Analysis

Asp18

Structure / Form

Disulfide-linked homodimer, labeled with Atto 647N via amines

Excitation wavelength: 647 nm

Emission wavelength: 667 nm

Predicted Molecular Mass

53.4 kDa (monomer)

SDS-PAGE

67-80 kDa, under reducing conditions

Product Datasheets

ATM1137

Product Datasheet

COA

SDS

ATM1137

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

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Fluorescent beads conjugated to anti-Human Siglec‑3/CD33 Monoclonal Antibody (MAB1137) were stained with (A) Recombinant Human Siglec‑3/CD33 Fc Chimeram Atto 647N Conjugate (Catalog # ATM1137) or (B) unstained.

SDS-PAGE

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2 μg/lane of Recombinant Human Siglec-3/CD33 Protein, Atto 647N Conjugate (Catalog # ATM1137) 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.

Reconstitution Calculator

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

Crocker, P.R. and A. Varki (2001) Trends Immunol. 22:337.
Crocker, P.R. and A. Varki (2001) Immunology 103:137.
Angata, T. et al. (2002) J. Biol. Chem. 277:24466.
Simmons, D. and B. Seed (1988) J. Immunol. 141:2797.
Freeman, S.D. et al. (1995) Blood 85:2002.
Taylor, V.C. et al. (1999) J. Biol. Chem. 274:11505.
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