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Your Position: Home > Protein > Siglec-3 > CD3-H82E7

Biotinylated Human Siglec-3 / CD33 Protein, Avitag™,His Tag (MALS verified)

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  • Synonym
    CD33,SIGLEC3,gp67
  • Source
    Biotinylated Human Siglec-3, Avitag,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 Structure

    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 when calibrated against Star Ribbon Pre-stained Protein Marker under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Labeling
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • Endotoxin
    Less than 0.1 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

    >90% as determined by SEC-MALS.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    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.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
Siglec-3 SDS-PAGE

Biotinylated Human Siglec-3, Avitag,His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).

SEC-MALS
Siglec-3 MALS images

The purity of Biotinylated Human Siglec-3, Avitag,His Tag (Cat. No. CD3-H82E7) is more than 90% and the molecular weight of this protein is around 55-75 kDa verified by SEC-MALS.

Bioactivity-ELISA
 Siglec-3 ELISA

Immobilized anti human CD33 MAb, Mouse IgG1 (Clone # 6C5/2) at 2 μg/mL (100 μL/well) can bind Biotinylated Human Siglec-3, Avitag,His Tag (Cat. No. CD3-H82E7) with a linear range of 0.078-2.5 ng/mL (QC tested).

Bioactivity-FACS
 Siglec-3 FACS

2e5 of anti-Siglec-3 CAR-293 cells were stained with 100 μL of 3 μg/mL of Human Siglec-3, Avitag,His Tag (Cat. No. CD3-H82E7) and negative control protein respectively, washed and then followed by PE-SA and analyzed with FACS (Routinely tested).

  • Background
    Myeloid cell surface antigen CD33 is also known as SIGLEC3, Siglecs (sialic acid binding Ig­like 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.
  • Clinical and Translational Updates

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