Human CD73/NT5E PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human CD73 in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).

spezie

Human

Cross Reaction

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

detection target

human CD73
Protein name: 5'-nucleotidase
Gene Full Name: 5'-nucleotidase, ecto (CD73)
abbreviation: NT5E
Background: 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.
ID:P21589
ID:4907

alias

5' NT|5' nucleotidase (CD73)|5' nucleotidase precursor|5' nucleotidase, ecto|5' nucleotidase, ecto (CD73)|5'-NT|5'-nucleotidase| 5NTD_HUMAN|CD73|CD73 antigen|E5NT|Ecto 5' nucleotidase|Ecto-5'-nucleotidase|eN|eNT|NT|NT5|NT5E|NTE|Purine 5 Prime Nucleotidase

Origin

Nativ and recombinant human CD73

Antibodies

a monoclonal antibody from mouse|a biotinylated polyclonal antibody from goat

Immunogene

CHO,W27-K547

sensitivity

<10pg/ml

detection range

31.2pg/ml-2000pg/ml

pack size

1x96 Wells

Reproducibility

|Intra-Assay Precision|Inter-Assay Precision
Sample|1|2|3|1|2|3
n|16|16|16|24|24|24
Mean(pg/ml)|132|510|1427|167|624|1553
Standard deviation|5.94|28.56|91.33|8.85|37.44|111.82
CV(%)|4.5|5.6|6.4|5.3|6|7.2

TMB(tetramethylbenzidine) incubation time

15-20

storage and Stability

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Principle of the Elisa

Boster′s human CD73 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD73 has been precoated onto 96-well plates. Standards(CHO, W27-K547) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD73 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human CD73 amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 2000pg/ml,1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml human CD73 standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, serum and plasma(heparin, EDTA) to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each human CD73 standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|31.2|62.5|125|250|500|1000|2000 O.D.|0.021|0.090|0.153|0.287|0.543|0.973|1.613|2.209

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human CD73 antibody|1
ST0000-10|Lyophilized recombinant human CD73 standard|10ng/tube�2
AR1107|Biotinylated anti- human CD73 antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(dilution 1:100)
AR1106-1|Sample diluent buffer|30ml
AR1106-2|Antibody diluent buffer|12ml
AR1106-3|ABC diluent buffer|12ml
AR1104|TMB color developing agent|10ml
AR1105|TMB stop solution|10ml
-|Adhesive cover|4