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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Proximity assays for sensitive quantification of proteins.

Christina Greenwood¹, David Ruff², Sara Kirvell¹

¹Postgraduate Medical Institute, Faculty of Medical Science, Anglia Ruskin University, Chelmsford, Essex CM1 1SQ, UK.

Biomolecular Detection and Quantification

|April 15, 2016

Summary

Proximity assays use DNA-tagged molecules to detect proteins and their interactions. These sensitive immunohistochemical tools offer accurate assessments for research and diagnostics.

Keywords:

Immuno-PCR Immunoassays In situ assays Proximity extension assay Proximity ligation assay

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Area of Science:

Biochemistry
Molecular Biology
Immunohistochemistry

Background:

Proximity assays are immunohistochemical tools employing DNA-tagged aptamers or antibodies.
These tools bind to proteins or protein complexes in close proximity.

Purpose of the Study:

To describe the principles and applications of proximity assays.
To highlight their potential in sensitive and specific protein detection.

Main Methods:

Utilizing DNA-tagged aptamers or antibodies for proximity binding.
Signal generation through amplification (PCR or isothermal) and probe hybridization.
Assays can be performed in homogeneous, solid-phase, or in situ formats.

Main Results:

Enables sensitive and robust detection of proteins, protein modifications, and protein-protein interactions.
In situ assays provide high spatial accuracy for visualizing single protein molecules or complexes.

Conclusions:

Proximity assays offer sensitive, specific, and accurate protein expression assessments.
Their properties make them valuable for research, diagnostics, and pharmacology.