Affinity Chromatography is a chromatography way which is applied for separating Biomacromolecule by using Biomacromolecule’s some homologous specific molecular recognition and reversible binding.
Glutathione-S-Transferase (GST) is an affinity tag which can enhance fusion protein solubility by tagging the eukaryotic protein. Besides, the bacterium with GST tagged protein has high level expression, but it might become inclusion body caused by albumin aggregation. We commonly add GST on the end of the target protein N or C.
GST has strong affinity to Glutathione, is means we can capture GST protein fusion on the fixed matrix (like Glutathione-bonded filler). This characteristic can be applied for Protein Purify and capturing protein.
Though GST tagged protein has high expression and easy to dissolving, the big tag size can interrupt downstream application protein functions. So we can cut the protein from GST tag to decrease the interruption.
Before using GST tag, we shall clone the target protein code on the carrier, then label the protein on Escherichia coli. Using inducible promoters control target protein expression can help to deal with protein toxic to bacterial cells. After bacterial cell lysis, we can use Glutathione-bonded filler to purify GST tagged protein. Then use protease to remove GST tag and do the next analysis.
GST tag has wide application rage, good specificity, it has different expressions on hosts, can be removed by different proteases, it can keep protein antigenicity and biological activity, improve Stability of foreign proteins. But what the shortcoming is big molecular weight, it might influence the protein functions and downstream experiments and it only can purify soluble protein.
Welch Materials GST Tanrose 4FF
glutathione affinity mediator
GST Tanrose 4FF is making highly cross-linked 4% ager gel as the matrix, it’s applied on industry large scale purification, it can purify the target protein in high flowing speed.
|Name||GST Tanrose 4FF glutathione affinity mediator|
|Matrix||Highly cross-linked 4% ager gel|
|Particle size range||45~165μm|
|Average particle size||90μm|
|Binding capacity||100mg (GST tagged protein) /mL (mediator)|
|Chemical stability||All the commen water solution, like 1M Acetate， pH4.0, 0.1M NaOH, 70% Ethanol, 8M Urea, 6M Guanidine hydrochloride (room temperature, one hour)|
|Suggesting highest speed||450cm/h|
|Storage solution||20% Ethanol|
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