Characterization of ion channels in endolysosomes by Patch clamp (CAT#: STEM-PET-0029-WXH)

Introduction

Endolysosomes are involved in a wide range of cellular processes including membrane trafficking, catabolism of extracellular and intracellular components, immune responses and antigen presentation, cell secretions, and cell life and death
The endolysosomal ion channels regulate the function of the endolysosomal system through four basic mechanisms: calcium release, control of membrane potential, pH change, and osmolarity regulation.

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Principle Applications Procedure Materials Notes Related Products

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.

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.

Materials

Patch clamp system

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