The journey from a target gene to purified protein The use of recombinant proteins has increased greatly in recent years, as has the wealth of techniques and products used for their amplification and purification. The advantages of using a recombinant tagged protein to facilitate purification and detection are now widely recognized. In general, the first step in a protein expression workflow involves cloning of the target gene into an appropriate expression vector. This is followed by the transformation of the expression vector into a suitable host system for subsequent expression analysis. Many host systems are available including bacteria, yeast, plants, filamentous fungi, insect or mammalian cells grown in culture, and transgenic animals or plants. Each host system has its own advantages and disadvantages, and it is important to consider these before the final selection of a host. After initial screening and identification of optimum expression conditions for your particular protein, you can scale-up the purification process to obtain large amounts of the target protein for downstream applications such as functional and structural studies. The purification of tagged proteins is relatively simple and saves time due to the high specificity between the tag on the expressed protein and the ligand on the affinity medium. You get high purity in a single step— affinity purification typically gives up to 95% purity. Further purification steps may be necessary if you require greater purity. Four of the most commonly used purification tags are, polyhistidines (His), glutathione S-Transferase (GST), Strep-tag II and Maltose Binding Protein (MBP).

Brochure

Pure Simplicity for tagged proteins pdf

Handbook

Recombinant Protein Purification Handbook: Principles and Methods pdf
Purifiying Challenging Proteins: Principles and Methods pdf