Intended Use
For the quantitative determination of Human ENTPD5 concentration in serum.
Contents of Kit
1. Human ENTPD5 Microplate - 96 well polystyrene microplate (12 strips of 8 wells) coated with rabbit mAb antibody against Human ENTPD5.
2. Human ENTPD5 Detecion Antibody - 0.2 mg/mL of rabbit mAb antibody against Human ENTPD5 conjugated to horseradish peroxidase (HRP) with preservatives.
3. Human ENTPD5 Standard - Recombinant Human ENTPD5 in a buffer with preservatives, lyophilized. The amount of standard is lot specific and indicated on the label of standard vial.
4. Wash Buffer Concentrate - 25 mL of a 20-fold concentrated solution of buffered surfactant with preservatives.
5. Dilution Buffer Concentrate - 8 mL of a 20-fold concentrated dilution buffer with preservatives.
6. Color Reagent A - 13 mL of stabilized hydrogen peroxide.
7. Color Reagent B - 13 mL of stabilized chromogen (tetramethylbenzidine).
8. Stop Solution - 8 mL of 2 N sulfuric acid.
Storage
Unopened Kit:
Store at 2 - 8°C and the kit is stable for 6 months upon receipt.
Opened/Reconstituted Reagents:
Diluted Wash Buffer and Diluted Dilution Buffer - Stored for up to 1 week at 2 - 8°C.
Conjugate, Stop Solution, Unmixed Color Reagent A and Unmixed Color Reagent B - Stored for up to 1 month at 2 - 8°C.
Standard - After reconstitution, store for up to 1 month at -80°C. The reconstituted standards should be aliquoted and avoid repeated freeze-thaw cycles.
Microplate Wells - Return unused strips to the foil pouch containing the desiccant pack and reseal along entire edge of zip-seal. Stored for up to 1 month at 2 - 8°C.
Precision
Intra-assay Precision (Precision within an assay)
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)
Three samples of known concentration were tested in five separate assays to assess inter-assay precision.
Detection Range
23.44-1500 pg/mL
Sensitivity
The minimum detectable dose (MDD) of Human ENTPD5 is typically less than 5.02 pg/mL. The MDD was determined by adding three standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration
General Description
Ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), also known as CD39 antigen-like 4, ER-UDPase, Guanosine-diphosphatase ENTPD5, Nucleoside diphosphatase Uridine-diphosphatase ENTPD5. This hydrolase is expressed in response to phosphoinositide 3-kinase (PI3K) signaling. Activation of PI3K results in FOXO phosphorylation by AKT1 and loss of ENTPD5 transcriptional repression. It is Up-regulated in PTEN-deficient cells. Uridine diphosphatase (UDPase) that promotes protein N-glycosylation and ATP level regulation.ENTPD5 promotes protein N-glycosylation and folding in the endoplasmic reticulum, as well as elevated ATP consumption in the cytosol via an ATP hydrolysis cycle. Together with CMPK1 and AK1, ENTPD5 constitutes an ATP hydrolysis cycle that converts ATP to AMP and results in a compensatory increase in aerobic glycolysis. ENTPD5 also hydrolyzes GDP and IDP but not any other nucleoside di-, mono- or triphosphates, nor thiamine pyrophosphate. This enzyme Plays a key role in the AKT1-PTEN signaling pathway by promoting glycolysis in proliferating cells in response to phosphoinositide 3-kinase (PI3K) signaling.
Standard Curve
This standard curve is only for demonstration purposes. A standard curve should be generated for each assay.
Citations
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