Quantitative Measurement of Ca2+ and Zn2+ in Mammalian Cells Using Genetically Encoded Fluorescent Biosensors

We describe important factors to consider when selecting a Ca2+ or Zn2+ biosensor, such as the sensor’s dissociation constant (K d′) and its dynamic range. We also discuss the limits of quantitative measurement using these sensors and reasons why a sensor may perform differently in different biological systems or subcellular compartments. We outline protocols for (1) quickly confirming sensor functionality in a new biological system, (2) calibrating a sensor to convert a sensor’s FRET ratio to ion concentration, and (3) titrating a sensor in living cells to obtain its K d′ under different experimental conditions.
Source: Springer protocols feed by Protein Science - Category: Biochemistry Source Type: news