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Study of Cyclic Nucleotide-Gated Channels by Patch clamp (CAT#: STEM-PET-0052-WXH)
Introduction
Cyclic nucleotide-gated (CNG) channels are ion channels which are activated by the binding of cGMP or cAMP. The channels are important cellular switches which transduce changes in intracellular concentrations of cyclic nucleotides into changes of the membrane potential and the Ca2+ concentration.
Principle
The patch-clamp technique involves a glass micropipette forming a tight gigaohm seal with the cell membrane. The micropipette contains a wire bathed in an electrolytic solution to conduct ions. To measure single ion channels, a “patch” of membrane is pulled away from the cell after forming a gigaohm seal.
Applications
• Study of ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
Procedure
1. Fabrication of glass electrodes
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
Materials
Patch clamp system
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