Adenosine Deaminase (ADA1) Inhibitor Screening Kit (Colorimetric) (BN01151)

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BN01151
€673

Description

ELISA Kit Technical ManualMSDS

Adenosine Deaminase (ADA1) Inhibitor Screening Kit (Colorimetric)

Adenosine Deaminase (ADA, E.C. 3.5.4.4.) is an enzyme that catalyzes the conversion of adenosine and 2'-deoxyadenosine to inosine and 2'-deoxyinosine.

Adenosine is a metabolite that plays important roles in determining cell proliferation, cell cycle progression, and immunosuppressive activation. Although multiple pathways exist for the production of adenosine, its deamination is irreversible, and becomes a critical point in purine metabolite signaling. Both adenosine and inosine bind the adenosine A2A receptor yet can trigger different downstream signaling. Excess adenosine in penile tissue contributes to the disorder called priapism, and impaired adenosine signaling is associated with erectile dysfunction. For this reason, substantial work has gone into identification of inhibitors that would be of clinical relevance. The major isoform of adenosine deaminase, ADA1, is widely expressed in most cells in humans, particularly in lymphocytes and macrophages. ADA1 is present in the cytosol, nucleus and found associated with dipeptidyl peptidase-4 on the cell membrane while ADA2 was first found in the spleen but is predominantly found in the plasma and serum. Adenosine signaling is part of the immune response, and has been shown to reduce T-cell activation. Adenosine Deaminase activity is thus a marker for T-lymphocyte proliferation. Assay Genie's Adenosine Deaminase (ADA1) Inhibitor Screening Kit includes the larger 41 kDa form, human ADA1. In Assay Genie's ADA inhibitor screening kit, adenosine is deaminated by ADA1 forming inosine. This intermediate is converted to uric acid by a series of enzymatic reactions. The formed uric acid can be measured in the supplied U.V. transparent plate at OD 293 nm. A highly specific ADA1 inhibitor is included as a positive control for inhibition. The kit provides a rapid, simple, sensitive, and reliable test, suitable for high-throughput screening of ADA1 inhibitors.

Figure: (a) Sample Inhibition of ADA1 enzyme activity by the supplied ADA Inhibitor. Reagent Background was subtracted for clarity. (b) Characteristic IC50 curves of relative inhibition as a function of inhibitor concentration. Curves generated using the ADA1 inhibitors Pentostatin (in blue) and erythro-9-(2-Hydroxy-3-nonyl)­adenine hydrochloride (EHNA) (orange). IC50 values obtained were 10.04 µM and 0.22 nM, respectively. Data were obtained by following the kit protocol.

Key Info Description

Product Code

BN01151

Size

100 assays

Detection Method

Absorbance (OD 293 nm)

Species Reactivity

Human

Applications

Screening of potential Adenosine Deaminase inhibitors

Features & Benefits

  • Rapid, and convenient
  • The kit screens potential human Adenosine Deaminase Inhibitors

Kit Components

  • ADA Assay Buffer (10X)
  • ADA Convertor
  • ADA Developer
  • ADA Substrate
  • ADA Enzyme
  • ADA Inhbitor (EHNA)
  • U.V. transparent plate (96-well)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

Usage

For Research Use Only! Not For Use in Humans.

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