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Related Concept Videos

Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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...

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Synthesis of Near-Infrared Emitting Gold Nanoclusters for Biological Applications
09:11

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Published on: March 22, 2020

Site-specific biomolecule labeling with gold clusters.

Christopher J Ackerson1, Richard D Powell, James F Hainfeld

  • 1Colorado State University, Department of Chemistry, Fort Collins, Colorado, USA.

Methods in Enzymology
|October 5, 2010
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Summary

Site-specific labeling of biomolecules using gold clusters improves electron cryomicroscopy data. This overview details methods for creating these gold-biomolecule conjugates for enhanced imaging.

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Area of Science:

  • Biochemistry
  • Nanotechnology
  • Microscopy

Background:

  • Electron cryomicroscopy (cryo-EM) is a powerful technique for visualizing biomolecular structures.
  • Enhancing the information content of cryo-EM data is crucial for detailed structural analysis.
  • Site-specific labeling offers a method to improve signal and localization in cryo-EM.

Purpose of the Study:

  • To provide a practical overview of techniques for site-specific labeling of biomolecules with gold clusters.
  • To detail established methods for forming biomolecule/gold cluster conjugates.
  • To explore different bioconjugation chemistries for gold cluster labeling.

Main Methods:

  • Overview of linker-mediated bioconjugation strategies.
  • Description of direct gold-biomolecule bonding techniques.
  • Explanation of coordination-mediated bonding using nickel(II) nitrilotriacetic acid (NTA) and polyhistidine (His)-tagged proteins.

Main Results:

  • Established techniques for forming biomolecule/gold cluster conjugates are presented.
  • Three distinct bioconjugation chemistries are covered in detail.
  • The methods facilitate the creation of labeled biomolecules for advanced microscopy.

Conclusions:

  • Site-specific labeling with gold clusters significantly enhances cryo-EM data quality.
  • The discussed bioconjugation techniques are practical and well-established.
  • These methods enable richer information extraction from biomolecular imaging experiments.