Human ENTPD5 ELISA Pair Set

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human ENTPD5 ELISA Pair Set is for the quantitative determination of Human ENTPD5. This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.

Contents of Kit

Bring all reagents to room temperature before use.
1. Capture Antibody – 0.5 mg/mL of rabbit anti-Human ENTPD5 monoclonal antibody (in PBS, pH 7.4). Dilute to a working concentration of 2 μg/mL in PBS before coating.
2. Detection Antibody – 0.2 mg/mL of rabbit anti-Human ENTPD5 monoclonal antibody conjugated to horseradish-peroxidase (HRP) (in PBS, 50 % HRP-Protector, pH 7.4, store at 4°C). Dilute to working concentration of 0.5 μg/mL in detection antibody dilution buffer before use.
3. Standard – Each vial contains 27 ng of recombinant Human ENTPD5. Reconstitute with 1 mL detection antibody dilution buffer. After reconstitution, store at -20°C to -80°C in a manual defrost freezer. A seven-point standard curve using 2-fold serial dilutions in sample dilution buffer, and a high standard of 1500 pg/mL is recommended.

Storage

Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Detection Antibody: Store at 4°C and protect it from prolonged exposure to light for up to 6 months from date of receipt. DO NOT FREEZE!
Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted standard at -80°C for up to 1 month. Avoid repeated freezethaw cycles

Sensitivity

The minimum detectable dose of Human ENTPD5 was determined to be approximately 23.4 pg/ml. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.

General Description

Ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), also known as CD39 antigen-like 4, ERUDPase, 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.