How does GST tagged protein purification work?
How does GST tagged protein purification work?
GST-tag protein purification is an affinity chromatography method. It exploits the high affinity of GST towards reduced Glutathione, its natural substrate. To achieve this Glutathione is bound to either agarose resin or magnetic beads, depending on the prefered purification method.
What is GST protein used for?
The GST tag Protein purification with affinity tags such as glutathione S-transferase (GST), histidine (HIS), and other affinity tags, enables purification of proteins with both known and unknown biochemical properties.
How do I add a GST tag to my protein?
The best way to add a GST-tag to a protein of interest is using a vector of the pGEX series. They also come with different protein cleavage sites. These are important for the later mentioned methods to remove a GST-tag.
What is GST fusion protein?
A gst fusion protein is a protein that is tagged with GST protein. The fusion protein is made by adding the sequence encoding GST to the plasmid expressing your protein of interest. GST has an affinity for GSH making them a good pair for use in chromatography and immunoprecipitation.
How is GST used as a tag?
GST Tag Definition Glutathione-S-transferase (GST), a 26 kDa sequence of 211 amino acids, is another widely used affinity tag that increases solubility of the desired protein. GST tag has affinity for immobilized glutathione and is used for prokaryotic expression more frequently.
What does GST stand for?
Goods and services tax (GST)
What is a recombinant fusion protein?
A recombinant fusion protein is a protein created through genetic engineering of a fusion gene. This typically involves removing the stop codon from a cDNA sequence coding for the first protein, then appending the cDNA sequence of the second protein in frame through ligation or overlap extension PCR.
How does fusion protein work?
A protein made from a fusion gene, which is created by joining parts of two different genes. Fusion genes may occur naturally in the body by transfer of DNA between chromosomes. For example, the BCR-ABL gene found in some types of leukemia is a fusion gene that makes the BCR-ABL fusion protein.