Measuring substrate binding and affinity of purified membrane transport proteins using the scintillation proximity assay

@article{Harder2012MeasuringSB,
  title={Measuring substrate binding and affinity of purified membrane transport proteins using the scintillation proximity assay},
  author={Daniel Harder and Dimitrios I. Fotiadis},
  journal={Nature Protocols},
  year={2012},
  volume={7},
  pages={1569-1578}
}
The scintillation proximity assay (SPA) is a rapid radioligand binding assay. Upon binding of radioactively labeled ligands (here L-[3H]arginine or D-[3H]glucose) to acceptor proteins immobilized on fluoromicrospheres (containing the scintillant), a light signal is stimulated and measured. The application of SPA to purified, detergent-solubilized membrane transport proteins allows substrate-binding properties to be assessed (e.g., substrate specificity and affinity), usually within 1 d. Notably… CONTINUE READING

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