Development of formaldehyde dehydrogenase-coupled assay and antibody-based assays for ALKBH5 activity evaluation

Sep 17, 2018Journal of pharmaceutical and biomedical analysis

Tests to measure ALKBH5 enzyme activity using formaldehyde detection and antibodies

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Abstract

Two assays for evaluating ALKBH5 activity were developed, achieving Z' factors of 0.74.

  • N6-methyladenosine is the most common internal modification found in eukaryotic messenger RNA.
  • ALKBH5 is a demethylase that influences mRNA export and RNA metabolism.
  • Previous attempts to identify modulators of ALKBH5 were hindered by a lack of suitable in vitro assays.
  • The newly developed assays utilize different principles to assess ALKBH5 activity.
  • Both assays are designed for high-throughput screening in a 384-well plate format.

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