다카라코리아바이오메디칼

Home > 전제품보기 > 단백질 발현 · 정제 > His-tag 융합 단백질 정제 > [개요] His-tag purification

[개요] His-tag purification

His Tag Purification 101

Learn how to choose 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 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
Small tag (6 amino acids) Works at C- or N-terminus Small tag is less likely to interfere with protein function Better for lower molecular weight proteins	Histidine-rich peptide (12 amino acids) Works at C- or N-terminus Long tag is less likely to internalize Better solubility More tag is exposed on the protein surface Better for high molecular weight proteins	Naturally occurring sequence (18 amino acids) Works at N-terminus Longer tag is even less likely to internalize Better solubility More tag is exposed on the protein surface Better for high molecular weight proteins Proteins containing this tag are more stable

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.