Recombinant Human Siglec-3/CD33 His-Tag Avi-Tag Protein, CF

Catalog #: AVI1137 Datasheet / COA / SDS

Biotinylated

Catalog #	Availability	Size / Price	Qty
AVI1137-050

Recombinant Human Siglec-3/CD33 His-Tag Avi-Tag Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Siglec-3/CD33 His-Tag Avi-Tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When Human Siglec-3/CD33 (6C5/2) Antibody (Catalog # MAB1137) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human Siglec-3/CD33 His-tag Avi-tag (Catalog # AVI1137) binds with an ED₅₀ of 60-360 pg/mL. Measured by the ability of the immobilized protein to support the adhesion of human red blood cells. Subramanian, S. et al. (2006) Blood 107:2548. The ED₅₀ for this effect is 0.05-0.4 μg/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived human Siglec-3/CD33 protein

Human Siglec-3/CD33 (Met17-His259) Accession # P20138.2	HHHHHH	Avi-tag
N-terminus		C-terminus

Accession #

P20138.2

N-terminal Sequence
Analysis

Met17

Structure / Form

Biotinylated via Avi-tag

Predicted Molecular Mass

29 kDa

SDS-PAGE

45-52 kDa, under reducing conditions

Product Datasheets

AVI1137

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI1137

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Human Siglec-3/CD33 (6C5/2) Antibody (Catalog # MAB1137) is immobilized at 0.5 µg/mL, 100 µL/well, Biotinylated Recombinant Human Siglec-3/CD33 His-tag Avi-tag (Catalog # AVI1137) binds with an ED₅₀ of 60-360 pg/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human Siglec-3/CD33 His-tag Avi-tag (Catalog # AVI1137) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 45-52 kDa.

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