膜杰作
Star Staining
分子别名（Synonym）
CD73,NT5E,5'-Nucleotidase,5'-NT,NT5,NTE
表达区间及表达系统（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
蛋白结构（Molecular Characterization）
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.
标记（Labeling）
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
蛋白标记度（Protein Ratio）
Passed as determined by the HABA assay / binding ELISA.
内毒素（Endotoxin）
Less than 1.0 EU per μg by the LAL method.
纯度（Purity）
>90% as determined by SDS-PAGE.
制剂（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.
重构方法（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.
存储（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:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
电泳（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%.
活性（Bioactivity）-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
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
背景（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.
