Gel Shift Assay
Gel Shift Assay - Related Videos
06:57
Electrophoretic Mobility Shift Assay (EMSA)
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2023
The electrophoretic mobility shift assay (EMSA) is a biochemical procedure used to elucidate binding between proteins and nucleic acids. In this assay a radiolabeled nucleic acid and test protein are mixed. Binding is determined via gel electrophoresis which separates components based on mass, charge, and conformation. This video shows the concepts of EMSA and a general procedure, including gel and protein preparation, binding, electrophoresis, and detection. Applications covered in this video...
04:31
Electrophoretic Mobility Shift Assay for Detection of Transcription Factor-DNA Complexes
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2025
In this video, we demonstrate the electrophoretic mobility shift assay to detect protein-DNA interactions using infrared fluorescent-labeled DNA probes. When the protein binds to the DNA probe, it forms a large complex that migrates slowly on and appears as a shifted band on the gel.
08:28
Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins
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Cited by 7 •
2020
Palmitoylation entails the incorporation of a 16-carbon palmitate moiety to cysteine residues of target proteins in a reversible manner. Here, we describe a biochemical approach, the acyl-PEGyl exchange gel shift (APEGS) assay, to investigate the palmitoylation state of any protein of interest in mouse brain lysates.
14:14
Optimized Fibrin Gel Bead Assay for the Study of Angiogenesis
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Cited by 166 •
2007
This video demonstrates the protocol of an in vitro angiogenesis assay that recapitulates several stages of angiogenesis. Time-lapse images of sprouting, lumen formation, branching and anastomosis - key features of angiogenesis - are shown.
08:48
Polyacrylamide Gels for Invadopodia and Traction Force Assays on Cancer Cells
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Cited by 9 •
2015
Mechanical rigidity in the tumor microenvironment plays a crucial role in driving malignant behavior by increasing invadopodia activity and actomyosin contractility. Using polyacrylamide gels (PAAs), invadopodia and traction force assays can be utilized to study the invasive and contractile properties of cancer cells in response to matrix rigidity.
FAQs
The complex moves more slowly because binding changes the migrating particle’s size and charge compared with the unbound DNA or RNA. In a native gel, these differences separate the free nucleic acid from the larger bound form, allowing researchers to distinguish whether a protein-associated species is present based on its altered migration pattern.
Distinct shifted bands indicate that a binding protein recognizes the tested nucleic acid under the assay conditions. Comparing the appearance or migration of bound and unbound forms provides evidence about interaction behavior, while the strength of the observed shift can help assess relative affinity. These results characterize whether binding is selective and how strongly it occurs.
A native gel preserves the nucleic acid–protein complex sufficiently for the bound and unbound forms to migrate as different species. If the interaction were not maintained during separation, the complex could no longer produce a distinct shifted band. Maintaining this condition is therefore essential for detecting the interaction through mobility differences rather than analyzing nucleic acid alone.
A typical workflow combines a labeled DNA or RNA molecule with the protein of interest, allows the interaction to form, and then separates the mixture in a native gel. The resulting migration pattern is examined for a shifted band relative to the free nucleic acid. This comparison indicates whether a detectable complex formed under the tested conditions.
In immunology, the assay can examine how transcription factors interact with nucleic acid sequences involved in gene control. Detecting altered binding patterns helps researchers investigate regulation during immune responses and identify changes in gene-control pathways. The approach therefore connects a molecular interaction with broader questions about how immune-related genes are regulated.
The method supports studies of interactions between pathogen or host nucleic acids and binding proteins. Such experiments can help characterize molecular events associated with infection and examine how infection changes gene-control pathways. By identifying interaction patterns linked to these processes, researchers can investigate infection mechanisms and explore potential therapeutic targets.