Principle of the Assay: ADAELISA kit applies the competitive enzyme immunoassay technique utilizing a polyclonal anti-ADAantibody and an ADA-HRP conjugate. The assay sample and buffer are incubated together with ADA-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the ADAconcentration since ADAfrom samples and ADA-HRP conjugate compete for the anti-ADAantibody binding site. Since the number of sites is limited, as more sites are occupied by ADAfrom the sample, fewer sites are left to bind ADA-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The ADAconcentration in each sample is interpolated from this standard curve.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes an enzyme that catalyzes the hydrolysis of adenosine to inosine. Various mutations have been described for this gene and have been linked to human diseases. Deficiency in this enzyme causes a form of severe combined immunodeficiency disease (SCID), in which there is dysfunction of both B and T lymphocytes with impaired cellular immunity and decreased production of immunoglobulins, whereas elevated levels of this enzyme have been associated with congenital hemolytic anemia. [provided by RefSeq, Jul 2008]
Uniprot Description
ADA: a enzyme that converts adenosine + H2O into inosine + NH3. Found in all tissues, occurs in large amounts in T-lymphocytes and, at the time of weaning, in gastrointestinal tissues. Genetic ADA deficiencies are a cause of autosomal recessive severe combined immuno-deficiency (SCID). Hereditary hemolytic anemia is caused by expression levels in erythrocytes 50-70 times greater than the norm.
Protein type: EC 3.5.4.4; Nucleotide Metabolism - purine; Hydrolase
Chromosomal Location of Human Ortholog: 20q13.12
Cellular Component: dendrite cytoplasm; extracellular space; cell surface; membrane; cell soma; lysosome; cytoplasm; plasma membrane; cell junction; cytosol; external side of plasma membrane
Molecular Function: adenosine deaminase activity; protein binding; zinc ion binding; purine nucleoside binding
Biological Process: negative regulation of circadian sleep/wake cycle, non-REM sleep; T cell activation; adenosine catabolic process; deoxyadenosine catabolic process; response to morphine; positive regulation of calcium-mediated signaling; purine salvage; positive regulation of T cell differentiation in the thymus; histamine secretion; purine ribonucleoside monophosphate biosynthetic process; regulation of cell-cell adhesion mediated by integrin; response to vitamin E; negative regulation of mature B cell apoptosis; positive regulation of T cell receptor signaling pathway; positive regulation of germinal center formation; positive regulation of B cell proliferation; hypoxanthine salvage; negative regulation of adenosine receptor signaling pathway; positive regulation of smooth muscle contraction; embryonic gut development; placenta development; aging; response to drug; Peyer's patch development; dATP catabolic process; nucleobase, nucleoside and nucleotide metabolic process; positive regulation of heart rate; liver development; negative regulation of leukocyte migration; purine base metabolic process; trophectodermal cell differentiation; purine nucleotide salvage; xanthine biosynthetic process; response to hydrogen peroxide; negative regulation of inflammatory response; response to hypoxia; inosine biosynthetic process; germinal center B cell differentiation; alveolus development; positive regulation of alpha-beta T cell differentiation
Disease: Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-negative, Nk Cell-negative, Due To Adenosine Deaminase Deficiency
Research Articles on ADA
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Product Notes
The Porcine ADA ada (Catalog #AAA735849) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA735849 ELISA Kit recognizes Porcine ADA. It is sometimes possible for the material contained within the vial of "Adenosine Deaminase, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
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