A simple and rapid radiochemical choline acetyltransferase (ChAT) assay screening test

  • Shiba K
  • Ogawa K
  • Kinuya S
 et al. 
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Abstract

A simple radiochemical choline acetyltransferase (ChAT) assay screening test was developed by measuring for [3H]acetylcholine ([3H]ACh) formed from 0.2 mM [3H]acetyl-coenzyme A ([3H]acetyl-CoA) and 1 mM choline by 0.2 mg of rat brain homogenates containing ChAT into 96-well microplates. A simple and rapid procedure for isolating [3H]ACh from the incubation mixture into 96-well microplates was achieved by using a sodium tetraphenylboron (Kalibor) solution (in ethyl acetate, 0.75%, w/v) and a hydrophobic liquid scintillator mixture (1:5, v/v, 0.2 mL) as an extraction solvent. The benefits of this radiochemical method using 96-well microplates are as follows: (1) this method is reliable and reproducible; (2) many samples can be examined at the same time by this method; (3) this method is economical and effective in reducing radioactive waste. The development of a new simple radiochemical ChAT assay screening test is the first stage of development of radiolabeled ChAT mapping agent. © 2006 Elsevier B.V. All rights reserved.

Author-supplied keywords

  • 96-Well microplate
  • Choline acetyltransferase
  • Radiochemical assay
  • [3H]acetyl-coenzyme A
  • [3H]acetylcholine

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