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PTEN Activity quantification in an ELISA format

Precisely measure PTEN phosphatase activity

Echelon Biosciences' PTEN Activity ELISA (cat nr. 117K-4700-1kit) is a clever, competitive ELISA for the precise measurement of PTEN phosphatase activity by simple detection of PIP2.

  • By Tbatan - Own work, CC BY-SA 3.0, https://en.wikipedia.org/w/index.php?curid=46233089No radioactivity
  • Organic solvent free
  • No thin layer chromatography

PTEN (Phosphatase and Tensin Homolog deleted on Chromosome 10) is a 3’ PI Phosphatase that converts PIP3 to PIP2 (and thus opposing PKB/Akt activation by PI 3-Kinase) and plays a central role in numerous cell signaling events and diseases

To measure PTEN Activity, most experimental procedures require the use of solvents, radioactivity and cumbersome thin layer chromatography.

With EBI’s PTEN Activity ELISA (cat. nr 117K-4700-1Kit), directly detect the phosphoinositide product, which compared to other assays, might detect free phosphate. This eliminates many possible sources of error since inorganic phosphate is the product of many phosphatase enzyme activities, and is found in common buffers and cleaning products.

Simple experimental workflow

The assay is a competitive ELISA in which the signal is inversely proportional to the amount of PI(4,5)P2 produced, hence a simple experimental workflow:

  1. After the PTEN reactions are complete, reaction products are added to the PI(4,5)P2-coated microplate and a PI(4,5)P2 detector protein is then added for competitive binding.
  2. A peroxidase-linked secondary detector and colorimetric detection is used to detect PI(4,5)P2 detector binding to the plate.
  3. The colorimetric signal is inversely proportional to the amount of PI(4,5)P2 produced by PTEN.

The assay can be used to measure recombinant or immunoprecipitated PTEN activity.

 

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Pathway Image by Tbatan - Own work, CC BY-SA 3.0, https://en.wikipedia.org/w/index.php?curid=46233089