Human ADA PicoKine ELISA Kit | ADA elisa kit

Human ADA PicoKine ELISA Kit

Cat. Num. AAA177572

• Reactivity: Human
• Synonyms: ADA PicoKine; Human ADA PicoKine ELISA Kit; Adenosine deaminase; ada; ADA1; ADA 1; Adenosine aminohydrolase; adenosine deaminase; ADA elisa kit

• Reactivity: Human
• Specificity: Natural and recombinant human ADA
• Sequence Length: 363

• Assay Range: 15.6pg/ml-1000pg/ml
• Sensitivity: <10pg/ml
• Sample: For quantitative detection of human ADA in cell culture supernates, serum, tissue homogenates and pleural fluid.
• Immunogen: sf21, A2-L363
• Assay Procedure Step 1: Aliquot 0.1ml per well of the 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml, 15.6pg/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. It is recommended that each human ADA standard solution and each sample be measured in duplicate.
• Intra-Assay Precision: Inter-Assay Precision; Sample|1|2|3|1|2|3; n|16|16|16|24|24|24; Mean(pg/ml)|55.6|386|741|60.7|405|810; Standard deviation|3.39|17.37|34.83|4.19|28.35|45.36; CV(%)|6.1|4.5|4.7|6.9|7.0|5.6
• Sample Data: Concentration(pg/ml)|0|15.6|31.2|62.5|125|250|500|1000; O.D.|0.051|0.118|0.196|0.299|0.574|0.995|1.753|2.449
• Preparation and Storage: Store at 4 degree C for 6 months, at -20 degree C for 12 months. Avoid multiple freeze-thaw cycles.

Typical Testing Data/Standard Curve (for reference only)

(Human ADA PicoKine ELISA Kit standard curve )

Related Product Information for ADA elisa kit

Principle of the assay: 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.

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.

NCBI and Uniprot Product Information

• NCBI GI #: 47078295
• NCBI GeneID: 100
• NCBI Accession #: NP_000013.2
• NCBI GenBank Nucleotide #: NM_000022.3
• UniProt Accession #: P00813; Q53F92; Q6LA59
• Molecular Weight: 40,764 Da
• NCBI Official Full Name: adenosine deaminase isoform 1
• NCBI Official Synonym Full Names: adenosine deaminase
• NCBI Official Symbol: ADA
• NCBI Protein Information: adenosine deaminase
• UniProt Protein Name: Adenosine deaminase
• Protein Family: Regulatory protein
• UniProt Gene Name: ADA
• UniProt Synonym Gene Names: ADA1
• UniProt Entry Name: ADA_HUMAN

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: Nucleotide Metabolism - purine; EC 3.5.4.4; Hydrolase

Chromosomal Location of Human Ortholog: 20q13.12

Cellular Component: cell junction; cell soma; cell surface; cytoplasm; cytosol; dendrite cytoplasm; external side of plasma membrane; extracellular space; lysosome; membrane; plasma membrane

Molecular Function: adenosine deaminase activity; protein binding; purine nucleoside binding; zinc ion binding

Biological Process: adenosine catabolic process; aging; alveolus development; dATP catabolic process; deoxyadenosine catabolic process; embryonic gut development; germinal center B cell differentiation; histamine secretion; hypoxanthine salvage; inosine biosynthetic process; liver development; negative regulation of adenosine receptor signaling pathway; negative regulation of circadian sleep/wake cycle, non-REM sleep; negative regulation of inflammatory response; negative regulation of leukocyte migration; negative regulation of mature B cell apoptosis; Peyer's patch development; placenta development; positive regulation of alpha-beta T cell differentiation; positive regulation of B cell proliferation; positive regulation of calcium-mediated signaling; positive regulation of germinal center formation; positive regulation of heart rate; positive regulation of smooth muscle contraction; positive regulation of T cell differentiation in the thymus; positive regulation of T cell receptor signaling pathway; purine nucleotide salvage; purine ribonucleoside monophosphate biosynthetic process; purine salvage; regulation of cell-cell adhesion mediated by integrin; response to drug; response to hydrogen peroxide; response to hypoxia; response to morphine; response to vitamin E; T cell activation; trophectodermal cell differentiation; xanthine biosynthetic process

Disease: Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-negative, Nk Cell-negative, Due To Adenosine Deaminase Deficiency