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Study of the electrical response of Zea mays protoplasts to auxins/antibodies by Patch clamp (CAT#: STEM-PET-0054-WXH)
Introduction
Protoplasts are isolated cells produced by removing the surrounding cell wall either by mechanical means or by the use of cell wall degrading enzymes. Removal of the cell wall leaves the protoplast surrounded by the plasmalemma membrane. Protoplasts can be used to study membrane biology, including the uptake of macromolecules and viruses. These are also used in somaclonal variation.
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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