An Electrochemiluminescence-Based Assay for MeCP2 Protein Variants

J Vis Exp. 2020 May 22:(159). doi: 10.3791/61054.

Abstract

The ECLIA is a versatile method which is able to quantify endogenous and recombinant protein amounts in a 96-well format. To demonstrate ECLIA efficiency, this assay was used to analyze intrinsic levels of MeCP2 in mouse brain tissue and the uptake of TAT-MeCP2 in human dermal fibroblasts. The MeCP2-ECLIA produces highly accurate and reproducible measurements with low intra- and inter-assay error. In summary, we developed a quantitative method for the evaluation of MeCP2 protein variants that can be utilized in high-throughput screens.

Publication types

  • Video-Audio Media

MeSH terms

  • Animals
  • Brain / metabolism*
  • Electrochemistry
  • Fibroblasts / metabolism
  • Luminescent Measurements*
  • Methyl-CpG-Binding Protein 2 / genetics
  • Methyl-CpG-Binding Protein 2 / metabolism*
  • Mice
  • Protein Transport

Substances

  • Methyl-CpG-Binding Protein 2