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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Jun 17, 2026

Live Cell Imaging of F-actin Dynamics via Fluorescent Speckle Microscopy (FSM)
19:16

Live Cell Imaging of F-actin Dynamics via Fluorescent Speckle Microscopy (FSM)

Published on: August 5, 2009

Recombinant alpha-actin for specific fluorescent labeling.

Atsuko H Iwane1, Masatoshi Morimatsu, Toshio Yanagida

  • 1Nanobiology Laboratories, Graduate School of Frontier Biosciences, Osaka University, Osaka 565-0871, Japan. iwane@phys1.med.osaka-u.ac.jp

Proceedings of the Japan Academy. Series B, Physical and Biological Sciences
|December 17, 2009
PubMed
Summary
This summary is machine-generated.

Actin

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Dynamics

Background:

  • Traditionally, actin was viewed as a passive component in actomyosin interactions.
  • Recent findings suggest actin possesses a more active role, evidenced by observed conformational changes.
  • Limitations in prior labeling techniques hindered definitive conclusions about actin's dynamic conformations.

Purpose of the Study:

  • To prepare fully active skeletal muscle alpha-actin using a baculovirus expression system.
  • To develop recombinant alpha-actin with specific sites for fluorescent probes in subdomains 1 and 2.
  • To enable advanced studies on actin's conformational dynamics and functional significance.

Main Methods:

  • Utilized a baculovirus expression system for alpha-actin production.
  • Engineered recombinant alpha-actin variants with targeted labeling sites.
  • Employed fluorescent probe labeling for specific subdomains (1 and 2).

Main Results:

  • Successfully prepared fully active alpha-actin.
  • Developed a novel method for specific fluorescent labeling of alpha-actin subdomains.
  • Established a foundation for more informative studies on actin conformation.

Conclusions:

  • The developed recombinant alpha-actin is fully active and amenable to specific labeling.
  • This advanced labeling technique significantly enhances the potential for acquiring detailed information from actin studies.
  • Future research can now explore the functional implications of different actin conformations with greater precision.