**His-Tag Protein Purification: An Efficient and Versatile Method**
His-tag purification is a widely used technique for isolating recombinant proteins, especially those expressed in prokaryotic systems like *E. coli*. This method relies on immobilized metal ion affinity chromatography (IMAC), where the His-tag binds to immobilized metal ions such as Ni²âº, Co²âº, or Cu²âº. The interaction between the histidine residues of the His-tag and the metal ions allows for selective purification of the target protein from complex mixtures.
The support material in IMAC can be agarose beads, magnetic particles, or pre-packed columns. These are typically functionalized with chelating agents like nitrilotriacetic acid (NTA) or iminodiacetic acid (IDA). NTA provides a more stable binding environment compared to IDA due to its higher coordination capacity. This stability makes NTA-based resins more suitable for high-throughput and large-scale purification processes.
One of the key advantages of using His-tag technology is that it is small and does not significantly affect the structure, solubility, or biological activity of the target protein. Additionally, His-tags are compatible with various expression systems and can be easily removed if needed. They also have low immunogenicity, making them ideal for producing antibodies in animals.
However, some challenges exist during His-tag purification. For instance, non-specific chelating agents present in bacterial lysates, such as citrate or other dicarboxylic acids, may interfere with the metal ion binding. To overcome this, it is essential to remove these chelators before starting the purification process.
PurKineâ„¢ His-Tag Purification Systems offer advanced solutions for efficient and reliable His-tag protein purification. These systems use innovative IMAC matrices that are compatible with reducing agents, chelators, and various buffer conditions. The products include different resin types, such as Ni-NTA, Ni-IDA, and Ni-Super Resin, each designed for specific applications.
The PurKineâ„¢ Ni-Super Resin is an improved version of traditional NTA resins. It offers enhanced stability, better compatibility with harsh chemicals, and higher flow rates, making it ideal for large-scale purifications. This resin is particularly effective when dealing with samples containing chelating agents that might otherwise cause nickel leaching.
In addition to resins, PurKineâ„¢ offers pre-packed columns and purification kits, providing convenience for researchers. The product line includes:
- **PurKineâ„¢ His-Tag Ni-IDA Resin**
- **PurKineâ„¢ His-Tag Ni-NTA Resin**
- **PurKineâ„¢ His-Tag Ni-NTA Packed Column**
- **PurKineâ„¢ His-Tag Protein Purification Kit (Ni-NTA)**
- **PurKineâ„¢ His-Tag Ni-Super Resin**
- **PurKineâ„¢ His-Tag Ni-Super Packed Column**
- **PurKineâ„¢ His-Tag Ni-Super Resin 6FF**
These products are available in various formats, including 5ml, 10ml, 50ml, and 100ml sizes, allowing flexibility for different experimental scales.
Whether you're working with inclusion bodies or soluble proteins, the PurKineâ„¢ system ensures high yield, purity, and reproducibility. Its robust formulation and versatility make it a preferred choice for researchers involved in protein expression and purification.
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