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Your Position: 홈 > Protein > CD73 > CD3-H82E3

Biotinylated Human CD73 / NT5E Protein, His,Avitag™

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    CD73,NT5E,5'-Nucleotidase,5'-NT,NT5,NTE
  • Source
    Biotinylated Human CD73, His,Avitag(CD3-H82E3) is expressed from human 293 cells (HEK293). It contains AA Trp 27 - Ser 549 (Accession # P21589-1).
    Predicted N-terminus: Trp 27
    Request for sequence
  • Molecular Characterization
    CD73 Structure

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

    The protein has a calculated MW of 61.7 kDa. The protein migrates as 60 kDa 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 1.0 EU per μg by the LAL method.
  • Purity

    >90% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in 20 mM Tris, 120 mM NaCl, pH7.5 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
CD73 SDS-PAGE

Biotinylated Human CD73, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-ELISA
CD73 ELISA

Immobilized Monoclonal Anti-Human CD73 Antibody, Human IgG1 at 5 μg/mL (100 μL/well) can bind Biotinylated Human CD73, His,Avitag (Cat. No. CD3-H82E3) with a linear range of 0.6-5 ng/mL (QC tested).

Protocol
CD73 ELISA

Immobilized Biotinylated Human CD73, His,Avitag (Cat. No. CD3-H82E3) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD73 Antibody, Human IgG1 with a linear range of 0.3-2 ng/mL (Routinely tested).

Protocol
  • Background
    5'-nucleotidase (5'-NT), also known as ecto-5'-nucleotidase or CD73 (cluster of differentiation 73), is an enzyme that is encoded by the NT5E gene. CD73 commonly serves to convert AMP to adenosine. Ecto-5-prime-nucleotidase (5-prime-ribonucleotide phosphohydrolase) catalyzes the conversion at neutral pH of purine 5-prime mononucleotides to nucleosides, the preferred substrate being AMP. Other forms of 5-prime nucleotidase exist in the cytoplasm and lysosomes and can be distinguished from ecto-NT5 by their substrate affinities, requirement for divalent magnesium ion, activation by ATP, and inhibition by inorganic phosphate. Rare allelic variants are associated with a syndrome of adult-onset calcification of joints and arteries (CALJA) affecting the iliac, femoral, and tibial arteries reducing circulation in the legs and the joints of the hands and feet causing pain.
  • Clinical and Translational Updates

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