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™ 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™ 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 plateLyophilized recombinant Human ADA standard, 2x10ng/tubeBiotinylated anti-Human ADA antibody, 1x130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100)Sample diluent buffer, 1x30mlAntibody diluent buffer, 1x12mlABC diluent buffer, 1x12mlTMB color developing agent, 1x10mlTMB stop solution, 1x10mlPBS washing buffer Powder for 1000ml, 1x buffer
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
