Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine®

Product Name

Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine®

SKU/Catalog Number

EK1422

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

NT5E (CD73; ecto-5'-nucleotidase) is a GPI-anchored ectoenzyme that catalyzes extracellular conversion of purine 5'-mononucleotides to nucleosides (preferred substrate AMP); described as a dimer of two identical ~70 kDa subunits and used as a marker of lymphocyte differentiation; page notes NT5 deficiency in immunodeficiency diseases and rare allelic variants linked to adult-onset calcification of joints and arteries (CALJA). Assay context: PicoKine® quantitative ELISA for human NT5E in cell culture supernatants, cell lysates, serum, and plasma (heparin/EDTA); sensitivity <10 pg/mL (listed as 10 pg/mL); assay range 156–10,000 pg/mL; CHO-expressed standard (W27–K547); sheep polyclonal capture/detection with avidin–biotin–peroxidase complex; no detectable cross-reactivity reported. Often contextualized in immune/inflammation panels with chemokine targets like CXCL10 (IP-10) or IL-8 (putative).

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1422)

Clonality of Antibodies

See Datasheet for details

Standard Protein

Expression system for standard: CHO; Immunogen sequence: W27-K547

Capture/Detection Antibodies

The capture antibody is polyclonal antibody from sheep and the detection antibody is polyclonal antibody from sheep.

Sensitivity

<10 pg/ml

Assay Range

156 pg/ml - 10,000 pg/ml

Standard Dilution Instructions

See datasheet of EK1422 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK1422 is reactive to NT5E in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK1422 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of NT5E

5'-nucleotidase (5'-NT), also known as ecto-5'-nucleotidase or CD73 (Cluster of Differentiation 73), is an enzyme that in humans is encoded by the NT5E gene. Ecto-5-prime-nucleotidase catalyzes the conversion at neutral pH of purine 5-prime mononucleotides to nucleosides, the preferred substrate being AMP. The enzyme consists of a dimer of 2 identical 70-kD subunits bound by a glycosyl phosphatidyl inositol linkage to the external face of the plasma membrane. And the enzyme is used as a marker of lymphocyte differentiation. Consequently, a deficiency of NT5 occurs in a variety of immunodeficiency diseases. 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. Rareallelic 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.

Kit Components

Catalog Number	Description	Quantity
EK1422-CAP	Anti-Human NT5E Pre-coated 96-well strip microplate	1
EK1422-ST	Human NT5E Standard	2 vials, 10 ng/tube
EK1422-DA	Human NT5E Biotinylated antibody (100x)	100ul
AR1103	Avidin-Biotin-Peroxidase Complex (100x)	100ul
AR1106-1	Sample Diluent	30ml
AR1106-2	Antibody Diluent	12ml
AR1106-3	Avidin-Biotin-Peroxidase Diluent	12ml
AR1104	Color Developing Reagent (TMB)	10ml
AR1105	Stop Solution	10ml
AR1106-5	Wash Buffer (25x)	20ml
PLA-SEA	Adhesive plate sealers	4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

• Microplate Reader capable of reading absorbance at 450nm.
• Incubator.
• Automated plate washer (optional).
• Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
• Multichannel pipettes are recommended for large amount of samples.
• Deionized or distilled water.
• 500ml graduated cylinders.
• Test tubes for dilution.

Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)	0	156	312	625	1250	2500	5000	10000
O.D.	0.056	0.099	0.132	0.201	0.348	0.6254	1.135	1.946

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect NT5E, Dilution ratio of 1:10, concentration in serum and plasma is around 6.4 ng/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

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