To determine the metal ion specificity of the SelFS
protein, various metals were used in response to the change in the
emission ratio (540/485 nm) on a microplate reader. The FRET ratio
was recorded after adding selected metals using 180 μL of the
SelFS protein (diluted in 20 mM PBS buffer pH 8.0) with 20 μL
of selenium, silver, iron, tungsten, and arsenic metals in the nanomolar-to-micromolar
range, 10 nM, 100 nM, 500 nM, 1 μM, 5 μM, and 10 μM.
To perform the competitive experiments, the FRET (540/485 nm) ratio
was recorded by adding the interferents such as Ca2+, Mg2+, Na+, and K+ to the SelFS protein
in the presence of 5 μM selenium. The FRET ratio was detected
after mixing 20 μL of these metals to 160 μL of the diluted
SelFS protein with 20 μL selenium on a microplate reader.
protein, various metals were used in response to the change in the
emission ratio (540/485 nm) on a microplate reader. The FRET ratio
was recorded after adding selected metals using 180 μL of the
SelFS protein (diluted in 20 mM PBS buffer pH 8.0) with 20 μL
of selenium, silver, iron, tungsten, and arsenic metals in the nanomolar-to-micromolar
range, 10 nM, 100 nM, 500 nM, 1 μM, 5 μM, and 10 μM.
To perform the competitive experiments, the FRET (540/485 nm) ratio
was recorded by adding the interferents such as Ca2+, Mg2+, Na+, and K+ to the SelFS protein
in the presence of 5 μM selenium. The FRET ratio was detected
after mixing 20 μL of these metals to 160 μL of the diluted
SelFS protein with 20 μL selenium on a microplate reader.
