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Preparation of gold probes

J E Beesley1

  • 1Wellcome Research Laboratories, Beckenham, UK.

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2012
PubMed

Abstract:

Colloidal gold probes are widely used in the biological sciences for both light and electron microscopy. A gold probe is an electron dense sphere of gold coated with an immunologically active protein. There are two aspects to be considered when making the probe.

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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.
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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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