Protein purification strategy| GST tag protein purification
In the process of protein purification experiment, it is critical to ensure the purity and quality of protein samples. In order to promote protein solubility, different tags will be used. In addition to the commonly used His tag, GST tag is also the star tag in purification experiment.
About GST Tag Protein and Purification
GST tag is a kind of affinity tag, which can specifically bind to glutathione. It shows the action principle of enzyme and substrate. Using this principle, GST tag is expressed into fusion protein, which is specifically combined with affinity medium with glutathione to purify the target protein. Then GST tag is cut off by specific enzyme to obtain natural protein.
GST fusion protein purification has the advantages of high purity, mild purification conditions, protein activity is not affected and soluble expression of protein is promoted. Therefore, GST fusion expression system should be given priority in protein research in order to obtain high soluble protein.
GST Tag Protein Purification Protocol
GST-tagged proteins from various sources, whether naturally or recombinantly expressed in E. coli or other host cells, can be purified by immobilized glutathione affinity chromatography and then competitively eluted with excess reduced glutathione. Using GST Chromrose 4FF medium with agarose gel as matrix, purification of target protein can be achieved at relatively high flow rate. This medium binds to the ligand via the tag, the buffer elutes to remove impurities, and then the tag protein is eluted under mild and non-denaturing conditions, so that the antigenic properties and functions of the protein are preserved.
product testing
Chromatographic medium: GST Chrorose 4FF
Sample: GST-tagged fusion protein
Experimental equipment: 5mL Xtrap GST4FF prepacked column, protein purifier
Equilibration/Wash Solution: 140mM NaCl, 2.7 mM KCl, 10mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4
Eluate: 140mM NaCl, 2.7 mM KCl, 10mM Na2HPO4, 1.8 mM KH2PO4, 10mM reduced glutathione, pH 7.4 (ready to use)
Description:
1, 3, 4, 5 are GST-tagged protein purification fillers from other companies
2 is GST Chromrose of Konossi Bio? 4FF
According to the test results, under the same experimental conditions, the GST Chrorose 4FF product load performance of Kono Biotech is outstanding.
product introduction
GST Chrorose 4FF is an affinity chromatography medium based on highly crosslinked agarose gel. It can purify recombinant derivatives of glutathione-S-transferase, glutathione-dependent protein and glutathione-transferase expressed by various expression systems in one step under mild operating conditions and strong characteristics.
