Gst Fusion
Gst Fusion - Related Videos
11:28
Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein (GST-RhoA(G17A)) from Epithelial Cell Lysates
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Cited by 14 •
2012
The method presented here describes an assay to follow activation of RhoA specific GDP/GTP Exchange Factors (GEFs) in cultured cells by making use of a mutant RhoA GST fusion protein that has high affinity for activated GEFs. GEFs are precipitated from cell lysates, detected by Western blotting and quantified by densitometry.
11:22
GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
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Cited by 18 •
2013
In the present protocol, we demonstrate a highly efficient and cost-effective small-scale protein purification method, which allows purification of recombinant proteins by uniquely combining a cleavable GST-tag and a small His-tag.
02:45
Nuclear Fusion
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2020
The process of converting very light nuclei into heavier nuclei is also accompanied by the conversion of mass into large amounts of energy, a process called fusion. The principal source of energy in the sun is a net fusion reaction in which four hydrogen nuclei fuse and ultimately produce one helium nucleus and two positrons. A helium nucleus has a mass that is 0.7% less than that of four hydrogen nuclei; this lost mass is converted into energy during the fusion. This reaction produces about...
09:19
Analysis of SNARE-mediated Membrane Fusion Using an Enzymatic Cell Fusion Assay
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2012
We have developed a cell fusion assay that quantifies SNARE-mediated membrane fusion events by activated expression of β-galactosidase.
06:10
Live Imaging of Mouse Secondary Palate Fusion
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Cited by 11 •
2017
Here, we present a protocol for live imaging of mouse secondary palate fusion using confocal microscopy. This protocol can be used in combination with a variety of fluorescent reporter mouse lines, and with pathway inhibitors for mechanistic insight. This protocol can be adapted for live imaging in other developmental systems.
FAQs
The GST component can improve the solubility of the linked protein during expression. This matters because a more soluble fusion product can be recovered in a form suitable for subsequent purification and analysis. In biomedical research, that property supports preparation of proteins whose biochemical behavior, structure, or disease-related interactions must be examined experimentally.
Selective purification depends on the GST portion recognizing immobilized glutathione. When a cell-derived protein mixture passes through the affinity material, the fusion protein can bind specifically while other components are separated from it. This binding step provides a focused route to enrich the protein of interest before it is used in biochemical or biomedical studies.
Tag removal allows investigators to study the protein of interest without the attached GST component when the tag could affect interpretation or downstream use. Cleavage methods are applied after purification to separate the two parts of the fusion protein. Researchers can therefore choose between analyzing the intact fusion or the released target protein, depending on the experiment.
A typical workflow begins by genetically linking GST to the selected protein, followed by expression of the recombinant construct. The produced fusion protein is then captured through its binding to immobilized glutathione during affinity chromatography. If needed, a cleavage step removes GST, leaving material prepared for biochemical assays, structural studies, or other research applications.
GST fusion is useful when investigators need purified antigens, enzymes, receptors, or other proteins for controlled laboratory experiments. The approach can support antibody development, biochemical assays, structural studies, and analyses of molecular interactions associated with disease. Its value lies in combining a solubility-supporting tag with a specific purification handle for research protein preparation.
Purified fusion proteins provide defined material for examining protein function and molecular behavior in experimental systems. Depending on the target, researchers may use them as antigens, assay components, structural-study material, or tools for investigating disease-related molecular interactions. Removing GST when required can further align the preparation with the analytical goals of the study.