For isolation of His-tagged proteins by affinity chromatography.
Affinity chromatography colums/nickel charged, IMAC columns
ADR 3 III • WGK 1
Prepacked columns with Nickel charged IDA-agarose bead matrix for low pressure affinity chromatography.
Suitable for all general applications, or if proteins of very small amount shall be isolated without loss.
- Fast and simple purification of His-tagged proteins from total lysates
- Superior recovery rate of pure His-tagged proteins
- Binding capacity (purified 6xHis protein) of 117 mg per ml matrix
- Easy elution and regeneration
- Very low Nickel leaching
- Compatible with denaturing and reducing reagents
- For gravity flow chromatography
Immobilized metal ion affinity chromatography (IMAC) still is the most widely used method to obtain high yield of very pure proteins with reasonable effort. Agarose His/Ni Columns offer the perfect solution for fast and simple isolation of His-tagged proteins from total lysates.
Figure: SDS-PAGE of 6xHis-fuculose aldolase, 28 kDa (arrows).
Very high yield of well purified proteins.
1: Marker, 2: Protein raw extract, 3,4: 6xHis target protein purified with Agarose His/Ni Beads or Column, respectively, 5: 6xHis target protein purified by competitor product.
Agarose His/Ni Columns are prepacked and may be used directly. The matrix consists of cross-linked and beaded 6 % agarose, IDA-conjugated and charged with divalent Nickel ions. Nickel chelates recognize two exposed histidines with good specificity and very high affinity, making the Ni2+ charged matrix the first choice for all standard applications. Agarose His/Ni Columns give very high yields of pure His-tagged protein with considerably low metal contamination. The tridentate IDA cross-linker provides easy elution with low amounts of imidazole. Additionally, Agarose Columns may repeatedly be regenerated, making them very cost-effective.
The Matrix is stable in all commonly used reagents including denaturing and reducing reagents (like 8 M urea, 6 M guanidinium hydrochloride, 5 mM DTT). Slurry in 20 % ethanol. Column material made from polypropylene and polyethylene (frit). May be autoclaved at 121 °C for 30 mins.