His-tag Purification Resins

• Table. His-tag protein purification selection guide

  Product Name	Ligand	Matrix	Dynamic Binding Capacity	Cat. No	Applications & Features
  High Affinity Ni-TED Resin FF	TED	6% agarose	10mg/ml	L00885 New	Suitable for purification using gravity columns and automated systems. High purity and great resistance to chemical reagents.
  High Affinity Ni-Charged Resin FF	NTA	6% agarose	≥50mg/ml	L00666 Hot	Suitable for purification using gravity columns and automated systems. High purity, high binding capacity, and Cost-effective.
  Ni-NTA Resin	NTA	4% agarose	>20mg/ml	L00250	Suitable for purification using gravity columns. High purity and high binding capacity.
  Ni Resin FF	IDA	6% agarose	>40mg/ml	L00465	Suitable for purification using gravity columns and automated systems. High binding capacity and cost-effective.
  Ni-IDA Resin	IDA	4% agarose	20mg/ml	L00223I	Suitable for purification using gravity columns. High binding capacity and cost-effective.

• Case 1. L00666 - High Binding Capacity, Strong Affinity, and High Purity

  High Affinity Ni-Charged Resin FF (L00666) based on a 6% highly cross-linked agarose matrix, covalently coupled with the NTA chelating ligand, which binds Ni²⁺ through four coordination sites. This resin enables high-affinity purification of recombinant proteins carrying polyhistidine (His) tags. High Affinity Ni-Charged Resin FF offers low Ni²⁺ leakage, high protein binding capacity, and excellent chemical stability, making it well-suited for various protein purification protocols. These features make Ni-NTA resin an ideal choice for purifying 6xHis-tagged fusion proteins.

  1. Experimental Conditions

  • Equipment: Peristaltic pump, glass chromatography column
  • Sample: Recombinant Protein A expressed in E. coli (target protein molecular weight: 34 kDa)
  • Sample volume: 21 L of E. coli lysate (from cell pellets harvested from two 70 L fermenters)
  • Column volume: Approx. 4.1 L (Diameter: 200 mm; Height: 13.1 cm)
  • Equilibration buffer: 60 mM NaH₂PO₄, 300 mM NaCl, 10 mM imidazole, pH 8.0
  • Flow rate: 500 mL/min; 5 column volumes
  • Sample loading buffer: 60 mM NaH₂PO₄, 300 mM NaCl, 10 mM imidazole, pH 8.0, Flow rate: 500 mL/min
  • Wash buffer: 60 mM NaH₂PO₄, 300 mM NaCl, 10–20 mM imidazole, pH 8.0, Flow rate: 700 mL/min, 10 column volumes
  • Elution buffer: 60 mM NaH₂PO₄, 300 mM NaCl, 250 mM imidazole, pH 8.0, Flow rate: 500 mL/min
  • Buffer exchange: Dialysis or ultrafiltration into 1× PBS

  2.Experimental Results

  

  Figure 1. SDS-PAGE Analysis of Purification Using L00666

  Ni-NTA resin demonstrates high binding capacity, strong affinity, and high purity in protein purification.

• Case 2. L00885 - Strong Resistance to Chemical Reagents

  High Affinity Ni-TED Resin FF (L00885) is a next-generation immobilized metal affinity chromatography (IMAC) resin. It features TED ligands coupled to a 6% agarose gel matrix, forming five coordination bonds with Ni²⁺ ions. This resin is primarily designed for the capture and purification of histidine-tagged (His-tag) proteins secreted into the supernatant of eukaryotic expression systems. Compared to other chelating resins, High Affinity Ni-TED Resin offers superior compatibility with a wider range of components, allowing efficient target protein purification even in the presence of certain concentrations of EDTA and DTT.

  

  The experimental results indicate that Ni-TED exhibits strong resistance to chemical reagents.