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Biotinylated Human Siglec-3 / CD33 Protein, Avi Tag (Avitag™)  pdf  pdf  pdf


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Synonym

CD33,SIGLEC3,gp67

Source

Biotinylated Human Siglec-3 / CD33, His Tag (CD3-H82E7) is expressed from human 293 cells (HEK293). It contains AA Asp 18 - His 259 (Accession # AAH28152.1).

Predicted N-terminus: Asp 18

Molecular Characterization

Siglec-3(Asp 18 - His 259)AAH28152.1
Avi
Poly-his

This protein carries an Avi tag (Avitag™) at the C-terminus, followed by a polyhistidine tag.

The protein has a calculated MW of 29.4 kDa. The protein migrates as 45-55 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

Biotinylation

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

Biotin:Protein Ratio

The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

 

Purity

>90% as determined by reduced SDS-PAGE.

Formulation

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.

Contact us for customized product form or formulation.

Reconstitution

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

Storage

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

No activity loss is observed after storage at:

  1. 4-8°C for 12 months in lyophilized state;
  2. -70°C for 3 months under sterile conditions after reconstitution.
 

SDS-PAGE

Biotinylated Human Siglec-3 / CD33, His Tag (Catalog # CD3-H82E7) SDS-PAGE gel

Biotinylated Human Siglec-3 / CD33, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity

Biotinylated Human Siglec-3 / CD33, His Tag (Catalog # CD3-H82E7) ELISA bioactivity

Immobilized anti human CD33 MAb, Mouse IgG1 (Clone # 6C5/2) at 2 μg/mL (100 μL/well) can bind Biotinylated Human Siglec-3, His Tag (Catalog # CD3-H82E7) with a linear range of 0.078-2.5 ng/mL.

Request for protocol

 

Background

Myeloid cell surface antigen CD33 also known as SIGLEC3, Siglecs (sialic acid binding Ig¬like lectins) and GP67, is a single-pass type I membrane protein which belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family. Human CD33 / 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. CD33 / Siglec-3 usually considered myeloid-specific, but it can also be found on some lymphoid cells. In the immune response, CD33 / Siglec-3 may act as an inhibitory receptor upon ligand induced tyrosine phosphorylation by recruiting cytoplasmic phosphatase(s) via their SH2 domain(s) that block signal transduction through dephosphorylation of signaling molecules. CD33 / Siglec-3 induces apoptosis in acute myeloid leukemia.

References

Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.


Limited Use License

The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.

Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.