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 aa 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. Sample Type: Cell culture supernates, serum. Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human ADA/Adenosine Deaminase. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human ADA/Adenosine Deaminase. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human ADA/Adenosine Deaminase BioAssay(TM) ELISA Kit was based on standard sandwich enzyme- linked immune-sorbent assay technology. A specific for ADA/Adenosine Deaminase 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 specific for ADA/Adenosine Deaminase 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/Adenosine Deaminase amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human ADA antibody, 1x plate Lyophilized recombinant Human ADA standard, 2x10ng/tube Biotinylated anti-Human ADA antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
