Abcam, ab204695, Adenosine Deaminase (ADA) Activity Assay Kit (Fluorometric)

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Adenosine Deaminase (ADA) Activity Assay Kit (Fluorometric) (ab204695) is an assay where inosine formed from the breakdown of adenosine is detected via a multi-step reaction, resulting in the formation of an intermediate that reacts with the ADA Probe to generate a fluorescent product that can be easily quantified at Ex/Em = 535/587 nm.
Key facts
Detection method:Fluorescent,
Sample types:Purified protein, Tissue Lysate, Cell culture media, Cell Lysate,
Assay type:Enzyme activity,
Sensitivity:= 10 µU,
Range:10 - 100 µU,
Assay Platform:Microplate reader

Product details:
Adenosine Deaminase (ADA) Activity Assay Kit (Fluorometric) (ab204695) is an assay where inosine formed from the breakdown of adenosine is detected via a multi-step reaction, resulting in the formation of an intermediate that reacts with the ADA Probe to generate a fluorescent product that can be easily quantified at Ex/Em = 535/587 nm. The kit measures total activity of Adenosine Deaminase with limit of quantification of 10 μU recombinant Adenosine Deaminase. The range of the assay is from 10-100 μU.
Other notes
This was previously called K328 Biovision Adenosine Deaminase Activity Assay Kit (Fluorometric). Biovision was acquired by Abcam in 2021.
Adenosine Deaminase (ADA, EC 3.5.4.4) is an enzyme that catalyses the conversion of adenosine and 2'-deoxyadenosine to inosine and 2'-deoxyinosine. Adenosine Deaminase is widely distributed in various tissues and cells. There are two isoforms, ADA1 and ADA2. ADA1 is widely expressed in most cells in the body, particularly in lymphocytes and macrophages. It is present in the cytosol, nucleus and has been found associated with DPP4/CD26 in the cell membrane. ADA2 was first found in the spleen but is predominantly found in the plasma and serum. Increased serum ADA levels are found in certain infectious diseases such as tuberculosis and various liver diseases such as acute hepatitis, alcoholic hepatic fibrosis, chronic active hepatitis to name a few. Adenosine Deaminase is also a marker for T-lymphocyte proliferation.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Adenosine deaminase (ADA) sometimes known as deaminase is an enzyme that plays a critical mechanical role in cellular metabolism. This enzyme catalyzes the deamination of adenosine converting it into inosine. ADA is a 41 kDa protein primarily expressed in most tissues including the thymus and spleen. Various ADA assays and ADA protocols exist to study its functionality and quantification often using ADA ELISA kits to measure ADA activity in biological samples. In cell lines such as HeLa ADA expression is also present and significant for studying its regulation and function.
Biological function summary
Adenosine deaminase activity influences purine metabolism playing an important role in the breakdown of adenosine. It is not part of a complex but interacts closely with other components of the purine salvage pathway to maintain nucleotide balance. This enzyme is important for the proper function of immune cells as it prevents toxic accumulations of adenosine which can be detrimental to lymphocyte proliferation and function. ADA's activity ensures the proper management of nucleotides within cells sustaining cellular health and functionality.
Pathways
Adenosine deaminase plays significant roles in the immune and purine catabolism pathways. Within these pathways ADA is closely related to proteins such as purine nucleoside phosphorylase (PNP) which together orchestrate the steps necessary for adenine and guanine nucleotide turnover. These proteins facilitate the conversion processes needed for cellular nucleic acid synthesis and energy regulation. By managing adenosine levels ADA's function ensures the smooth operation of cellular signaling and metabolic balance.
Adenosine deaminase deficiency is linked to severe combined immunodeficiency (SCID) and can also be associated with pulmonary disorders like asthma. A lack of ADA activity leads to the accumulation of toxic metabolites impairing immune function and contributing to the pathogenesis of SCID. In some patients with pulmonary disorders altered ADA activity impacts the inflammatory response. ADA's mechanistic interactions with other proteins such as PNP highlight its importance in maintaining immune competence and metabolic homeostasis emphasizing the enzyme's role in these clinical conditions.