Measuring SERCA-mediated calcium uptake in mouse muscle homogenates

Abstract

This protocol employs the indo-1 Ca²⁺ fluorophore to quantify Ca²⁺ uptake by the sarco(endo)plasmic reticulum Ca²⁺ ATPase pump in murine muscle homogenates and allows for real-time kinetic measurement of Ca²⁺ mobilization within the muscle homogenate. This protocol can be easily adapted for other tissue types and can be modified to single-emission/single-excitation Ca²⁺ dyes. Fitted to a 96-well plate, this assay can be readily performed in most laboratories with minimal sample requirement and the option of multiple replicates. For complete details on the use and execution of this protocol, please refer to Braun et al. (2022),¹ Braun et al. (2021a),² Braun et al. (2021b),³ Cleverdon et al. (2022),⁴ and Geromella et al. (2022).⁵.

Keywords: Cell Biology; Cell-based Assays; Metabolism.