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Study of second messenger-mediated cellular events by Patch clamp (CAT#: STEM-PET-0065-WXH)

Introduction

Second messengers are molecules that relay signals received at receptors on the cell surface — such as the arrival of protein hormones, growth factors, etc. — to target molecules in the cytosol and/or nucleus. Second messengers trigger physiological changes at cellular level such as proliferation, differentiation, migration, survival, apoptosis and depolarization. They are one of the triggers of intracellular signal transduction cascades.




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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