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ELISA Kit for Cluster Of Differentiation 73 (CD73)

NT5E; NT5-E; E5NT; E5-NT; NTE; EN; ENT; 5'-Nucleotidase, Ecto

Specificity

This assay has high sensitivity and excellent specificity for detection of Cluster Of Differentiation 73 (CD73).
No significant cross-reactivity or interference between Cluster Of Differentiation 73 (CD73) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cluster Of Differentiation 73 (CD73) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cluster Of Differentiation 73 (CD73) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Autoimmunity The expression of CD39 on regulatory T cells is genetically driven and further upregulated at sites of inflammation Pubmed:25640206
Talanta Simultaneous accurate quantification of HO-1, CD39, and CD73 in human calcified aortic valves using multiple enzyme digestion–filter aided sample pretreatment … 10.1016:j.talanta.2018.01.044
Cancer Biomarkers Expression and clinical significance of serum NT5E protein in patients with colorectal cancer Pubmed: 30932882
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