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

5& ELISA kit for Human

Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine™ (96 Tests). Quantitate Human NT5E in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1422
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

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

View all 5& ELISA kits

SKU/Catalog Number

EK1422

Size

96 wells/kit, with removable strips.

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

Description

Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine™ (96 Tests). Quantitate Human NT5E in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

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

Immunogen

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

Sensitivity

<10 pg/ml

Assay Range

156 pg/ml - 10,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank 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 5&

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.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015631262512502500500010000
O.D.0.0550.1250.1690.2990.5450.9561.4372.064

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect 5&, 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.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)2301941536122219124929
Standard deviation15.4179.58387.2918.64101.33359.81
CV (%)6.7%4.1%7.2%8.4%5.3%7.3%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of 5& in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 123023524326024211.374.6%
Sample 21941187018301891188340.012.1%
Sample 353614741545554255245293.245.5%
*number of samples for each test n=16.

Gene/Protein Information For NT5E (Source: Uniprot.Org, NCBI)

Gene Name

NT5E

Full Name

5'-nucleotidase

Weight

63.368kDa

Superfamily

5'-nucleotidase family

Alternative Names

5' nucleotidase (CD73); 5'-NT; 5-NT; 5'-Nucleotidase; 5'-nucleotidase, ecto (CD73); CD73 antigen; CD73; E5NT; EC 3.1.3.5; ecto-5'-nucleotidase; eN; eNT; NT; NT55'-nucleotidase; NT5E; NTE; Purine 5-Prime-Nucleotidase NT5E CALJA, CD73, E5NT, NT, NT5, NTE, eN, eNT 5-nucleotidase ecto 5-nucleotidase|5-NT|Purine 5-Prime-Nucleotidase|ecto-5-nucleotidase

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on NT5E, check out the NT5E Infographic

NT5E infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for NT5E: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

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13 Customer Q&As for Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine®

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring CD73 using the EK1422 Human CD73 Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2020-12-09

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to prevent protein degradation.

Boster Scientific Support

Answered: 2020-12-09

Question

Is the EK1422 recombinant CD73 supplied as the standard suitable for WB? Is it a full-length protein?

Verified customer

Asked: 2020-11-25

Answer

For the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422), recombinant CD73 supplied as the standard contains BSA and is unsuitable for WB. The immunogen of the standard is W27-K547.

Boster Scientific Support

Answered: 2020-11-26

Question

Does the sample diluent in EK1422 contain any phosphate?

Verified customer

Asked: 2020-10-07

Answer

Our lab technicians confirmed that there wasn't any phosphate in sample diluent for the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422).

Boster Scientific Support

Answered: 2020-10-07

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2020-09-27

Answer

A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2020-09-27

Question

Does this kit detect the dimer/active form of CD73 and recombinant protein?

Verified Customer

Verified customer

Asked: 2019-12-17

Answer

This kit is able to detect the dimer/active form of CD73.

Boster Scientific Support

Answered: 2019-12-17

Question

Q: how to analyze ELISA data? I have performed CD73 level in cell lysates.

Verified Customer

Verified customer

Asked: 2019-09-02

Answer

A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2019-09-02

Question

What is the recommended lysis buffer compatible with EK1422?

Verified customer

Asked: 2019-06-28

Answer

We recommend using IP lysis buffer (AR0107) for the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422).

Boster Scientific Support

Answered: 2019-06-30

Question

Is RIPA lysis Buffer suitable for EK1422 and is there any component that could interfere with the assay?

Verified customer

Asked: 2019-06-28

Answer

RIPA lysis Buffer isn't suitable for the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422). Sodium deoxycholate 1% and SDS 0.1% will denature the protein.

Boster Scientific Support

Answered: 2019-06-30

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

C. Dhawan

Verified customer

Asked: 2019-06-13

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2019-06-13

Question

What is the TMB incubation time for EK1422?

Verified Customer

Verified customer

Asked: 2019-02-05

Answer

The TMB incubation time for the current lot of the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422) is 15-25min. The incubation time in the datasheet is just for reference. The accurate incubation time should depend on the specific assay.

Boster Scientific Support

Answered: 2019-02-05

Question

Are there any matrix effects when running samples neat in this kit?

Verified Customer

Verified customer

Asked: 2018-10-22

Answer

There might be some matrix effects when running samples neat in the Human CD73/NT5E ELISA Kit PicoKine™ (EK1422). We suggest you to dilute the samples by referring to the COA.

Boster Scientific Support

Answered: 2018-10-22

Question

Q: how to thaw whole blood sample for CD73 ELISA after freezing?

M. Bhatt

Verified customer

Asked: 2018-06-12

Answer

A: we do not recommend freezing and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test CD73 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

Boster Scientific Support

Answered: 2018-06-12

Question

Q: can you suggest the dilution ratio of serum samples for detection of CD73 in Human cell lysates? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.

H. Perez

Verified customer

Asked: 2017-01-27

Answer

A: without knowing the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the CD73 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2017-01-27

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