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

Overview

  • Product name
    Adenosine Deaminase (ADA) Activity Assay Kit (Fluorometric)
    See all ADA kits
  • Detection method
    Fluorescent
  • Sample type
    Cell Lysate, Cell culture media, Purified protein, Tissue Lysate
  • Assay type
    Enzyme activity
  • Sensitivity
    10 µU
  • Range
    10 µU - 100 µU
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mammal
  • Product overview

    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.

  • Notes

    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.

  • Tested applications
    Suitable for: Functional Studiesmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    ADA Assay Buffer (10x) 1 x 25ml
    ADA Convertor 1 vial
    ADA Developer 1 vial
    ADA Positive Control 1 vial
    ADA Probe 1 x 200µl
    ADA Substrate 1 x 500µl
    Inosine Standard (10 mM) Yellow 1 x 100µl
  • Research areas
  • Function
    Catalyzes the hydrolytic deamination of adenosine and 2-deoxyadenosine. Plays an important role in purine metabolism and in adenosine homeostasis. Modulates signaling by extracellular adenosine, and so contributes indirectly to cellular signaling events. Acts as a positive regulator of T-cell coactivation, by binding DPP4. Its interaction with DPP4 regulates lymphocyte-epithelial cell adhesion.
  • Tissue specificity
    Found in all tissues, occurs in large amounts in T-lymphocytes and, at the time of weaning, in gastrointestinal tissues.
  • Involvement in disease
    Defects in ADA are the cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-negative due to adenosine deaminase deficiency (ADASCID) [MIM:102700]. SCID refers to a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients with SCID present in infancy with recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. ADA-SCID is an autosomal recessive form accounting for about 50% of non-X-linked SCIDs. ADA deficiency has been diagnosed in chronically ill teenagers and adults (late or adult onset). Population and newborn screening programs have also identified several healthy individuals with normal immunity who have partial ADA deficiency.
  • Sequence similarities
    Belongs to the adenosine and AMP deaminases family.
  • Cellular localization
    Cell membrane. Cell junction. Cytoplasmic vesicle lumen. Cytoplasm. Colocalized with DPP4 at the cell junction in lymphocyte-epithelial cell adhesion.
  • Information by UniProt
  • Alternative names
    • ada
    • ADA_HUMAN
    • ADA1
    • Adenosine aminohydrolase
    • Adenosine deaminase
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab204695 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

Images

  • Typical Inosine Standard Curve

  • Adenosine Deaminase Activity in Jurkat Cell (T-lymphocyte) lysate (105 ng) and Positive Control; BG: Background.

  • Adenosine Deaminase specific activity in Jurkat cell lysate (105 ng) and rat brain lysate (2 μg).

Protocols

References

ab204695 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abreviews
The kit can be used to detect adenosine deaminase activity in fly tissues. it also detect the effect of Adgf knockdown and overexpression in fly tissues. the attached figure is a representative results of three biological replicates. Gswitch is a ubiquitous gene expression driver that is active upon supplementation of the drug RU.


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Submitted Mar 14 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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