Human ADA PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human ADA in cell culture supernates, serum, tissue homogenates and pleural fluid.

spezie

Human

Cross Reaction

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

detection target

human ADA
Protein name: Adenosine deaminase
Gene Full Name: adenosine deaminase
abbreviation: ADA
Background: Adenosine Deaminase (also known as adenosine aminohydrolase, or ADA) is an enzyme involved in purine metabolism. It is needed for the breakdown of adenosine from food and for the turnover of nucleic acids in tissues. ADA in humans is involved in the development and maintenance of the immune system. However, ADA association has also been observed with epithelial cell differentiation, neurotransmission, and gestation maintenance. It has also been proposed that ADA, in addition to adenosine breakdown, stimulates release of excitatory amino acids and is necessary to the coupling of A1 adenosine receptors and heterotrimeric G proteins. Adenosine deaminase deficiency leads to pulmonary fibrosis, suggesting that chronic exposure to high levels of adenosine can exacerbate inflammation responses rather than suppressing them. It has also been recognized that adenosine deaminase protein and activity is upregulated in mouse hearts that overexpress HIF-1 alpha, which in part explains the attenuated levels of adenosine in HIF-1 alpha expressing hearts during ischemic stress.
ID:P00813
ID:100

alias

ada|ADA1|ADA 1|Adenosine aminohydrolase|Adenosine deaminase

Origin

Nativ and recombinant human ADA

Antibodies

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

Immunogene

sf21, A2-L363

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)|45|153|679|51|201|821
Standard deviation|2.47|7.49|34.62|1.98|8.44|37.76
CV(%)|5.5|4.9|5.1|3.9|4.2|4.6

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 ADA ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for ADA has been precoated onto 96-well plates. Standards(Expression system for standard: sf21, Immunogen sequence: A2-L363) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for ADA 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 ADA 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 ADA 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, tissue homogenates or pleural fluid to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each human ADA 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.|0.051|0.118|0.196|0.299|0.574|0.995|1.753|2.449

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human ADA antibody|1
ST0000-10|Lyophilized recombinant human ADA standard|10ng/tube�2
AR1107|Biotinylated anti- human ADA 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