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Enlightening G-protein-coupled receptors on the plasma membrane by super-resolution photoactivated localization microscopy (CAT#: STEM-MIT-0395-LJX)

Introduction

The G protein-coupled Receptors (GPCRs) are a family of membrane Protein receptors. It can participate in many physiological processes, such as behavior and emotion regulation, immune system regulation, autonomic nervous system regulation and so on.
The possibility to visualize and image the arrangement of proteins within the cell at the molecular level has always been an attraction for scientists in biological research. In particular, for signalling molecules such as GPCRs (G-protein-coupled receptors), the existence of protein aggregates such as oligomers or clusters has been the topic of extensive debate. One of the reasons for this lively argument is that the molecular size is below the diffraction-limited resolution of the conventional microscopy, precluding the direct visualization of protein super-structures. On the other hand, new super-resolution microscopy techniques, such as the PALM (photoactivated localization microscopy), allow the limit of the resolution power of conventional optics to be broken and the localization of single molecules to be determined with a precision of 10-20 nm, close to their molecular size. The application of super-resolution microscopy to study the spatial and temporal organization of GPCRs has brought new insights into receptor arrangement on the plasma membrane. Furthermore, the use of this powerful microscopy technique as a quantitative tool opens up the possibility for investigating and quantifying the number of molecules in biological assemblies and determining the protein stoichiometry in signalling complexes.




Principle

Principles of photoactivated localization microscopy: By fitting the two-dimensional Gaussian function to determine the centroid of microscope-formed light spots, a single fluorescent source (such as a fluorescent group) can be located with high precision. The accuracy of the calculation to determine the centroid depends only on the number of photons collected, and the resolution scale can be tens of nanometers or smaller. To achieve this accuracy, the density of the fluorescent molecules being tested is required to be low enough that the spots of the two fluorescent groups are unlikely to overlap.

Applications

Applied in many areas of the life sciences

Procedure

1. Sampling
2. Preparation of slices
3. Staining (Select according to the specific experimental situation)
4. Observation

Materials

• Sample Type:
G-protein-coupled receptors

Notes

Operate in strict accordance with the operating procedures, and shall not arbitrarily change the operating procedures
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