A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:05Title
- 01:56Pretreatment of Heterologous ATPase-expressing Oocytes
- 02:43Electrophysiology
- 04:53Rb+ Uptake Measurements
- 06:03Preparation of AAnalyst 800
- 07:46Results: Determination of Enzyme Kinetic Parameters
- 12:14Conclusion
We describe a method to quantify the activity of K+-countertransporting P-type ATPases by heterologous expression of the enzymes in Xenopus oocytes and measuring Rb+ or Li+ uptake into individual cells by atomic absorption spectrophotometry. The method is a sensitive and safe alternative to radioisotope flux experiments facilitating complex kinetic studies.
