Measuring the mechanical stability of immunoglobulin and fibronectin III domains in the muscle protein titin by atomic force microscopy (CAT#: STEM-SMMT-0026-LJX)

Introduction

The domains of the giant muscle protein titin (connectin) provide interaction sites for other sarcomeric proteins and fulfill mechanical functions. In this paper we compare the unfolding forces of defined regions of different titin isoforms by single-molecule force spectroscopy. Constructs comprising six to eight immunoglobulin (Ig) domains located in the mechanically active I-band part of titin are compared to those containing fibronectin III (Fn3) and Ig domains from the A-band part. The high spatial resolution of the atomic force microscope allows us to detect differences in length as low as a few amino acids. Thus constructs of different lengths may be used as molecular rulers for structural comparisons with other modular proteins.

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Principle Applications Procedure Materials Notes Related Products

Principle

Atomic force microscope (AFM) is a new type of surface analysis instrument based on the principles of physics and imaging through the interaction of scanning probe and sample surface atoms. It belongs to the third generation of microscopes after optical microscopes and electron microscopes.
AFM usually uses a sharp probe to scan the sample, which is fixed on a microcantilever that is extremely sensitive to the surface force between the probe and the sample. The deflection of the cantilever under force can cause the laser beam emitted by the laser source to shift after being reflected by the cantilever. The detector receives reflected light, and finally receives signals that are collected, processed, and formed into surface morphology images of the sample through a computer system.

Applications

Imaging of micro and nanoscale features on the surface of the sample
Suitable for research in fields such as materials science, biology, and chemistry

Procedure

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

Materials

• Sample Type:
Myocyte

Notes

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

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