- Mass spectrometry digestion reagents
- Why tag a protein?
- Tech note: cobalt resin
- Purification methods overview
- Simplified purification of active, secreted his-tagged proteins
- Overview: His60
- Tech note: Capturem technology
- Tech note: Capturem large volume
- Magnetic beads
- TALON resin selection guide
- Selection guide: His60 resin
- FAQs: TALON
- Antibody purification
- Other tag purification
- Phosphoprotein and glycoprotein purification
- Expression systems
His-tagged protein purification overview
His-tag purification 101
Learn how to identify the best his-tagged protein purification method for your specific application. Mini spin columns are ideal for simple, rapid purification of small amounts of protein. The high capacity of His60 Ni Resin makes it an excellent choice for purifying large amounts of high-quality protein, while the Co2+ core of TALON resin allows it to deliver proteins of the highest purity. Both of these resins are available in several formats, including gravity columns, FPLC cartridges, and various sizes of bulk resin, as well as magnetic beads for microscale purification.
|Comparison of his-tagged protein purification methods|
|Purity||Yield||Binding capacity (mg protein/ml matrix)||Start to finish (minutes)||Ease of use|
|Capturem His-Tagged Purification Miniprep Kit||****||0.1 mg/mini spin column||100 mg/ml||5 min||*****|
|His60 Ni||****||60 mg/ml resin||60 mg/ml||60–120 min||***|
|TALON||*****||20 mg/ml resin||20 mg/ml||60–120 min||***|
|Magnetic beads||*****||15 mg/ml resin||15 mg/ml||60 min||****|
What makes his tags attractive?
Over 75% of researchers use his tags for purifying proteins, because they offer the following advantages:
- Small size
- Less likely to alter wild-type protein function/structure
- Easy to clone
- Effective when incorporated on either the N- or C-terminus
- May be used to purify both native and denatured proteins
|Advantages of common his tags|
|6xHis tag||6xHN tag||HAT tag|
Choice of metal ions
Nickel- and cobalt-based resins offer a different balance of specificity vs. adsorption. The higher specificity of Co2+ provides greater purity, while the superior adsorption of Ni2+ provides a higher binding capacity.
Takara Bio offers four different technologies for his-tagged protein purification: Capturem membranes, Talon resin, His60 Ni resin, and magnetic beads:
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