High-throughput, milligram-scale purification of His•Tag® fusion proteins The RoboPop™ Ni-NTA His•Bind® Purification Kit is designed for filtration-based 96-well format purification of soluble His•Tag® fusion proteins directly from E. coli cultures without harvesting cells. The kit features PopCulture® Reagent, rLysozyme™ Solution, and Benzonase® Nuclease for centrifuge-free cell lysis and extract preparation in one step. The combination of PopCulture extraction, Ni-NTA His•Bind Resin, and a 2-ml filter plate allows high-throughput processing of up to 5 ml of E. coli culture per well. Whereas the magnetic-based His•Mag™ kit purifies up to 125 µg target protein per 1 ml culture, the filtration-based kit purifies up to 1 mg His•Tag fusion protein per 5 ml culture.
Bacterial culture, cell lysis, and resin binding steps are carried out in standard 24- well plates (not supplied), which accommodate a maximum volume of 5 ml per well. The reaction slurry is then transferred to a 96-well Filter Plate (included) and the washing and elution steps are carried out on a vacuum manifold. The Filter Plate is compatible with standard filter manifolds for manual sample processing, and the entire purification has been validated for robotic sample processing with the Genesis® Freedom™ Workstation from Tecan and the MultiPROBE® II liquid handling work station from PerkinElmer Life Sciences. A 96-well Collection Plate (1-ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins.
 |  |
 Features and Benefits:
- High-throughput, 96-well protein purification directly from E. coli without harvesting cells
- Centrifuge-free cell lysis and extract preparation in one step
- Large-scale culture processing—up to 5 ml per well
- High protein yield—up to 1 mg per well
- Validated for robotic sample processing with the Genesis® Freedom™ Workstation from Tecan and the Packard-brand MultiPROBE® II liquid handling work station from PerkinElmer Life Sciences
References:
Grabski, A., et al. 2002. inNovations 14, 2. Components:
| • | 75 ml | PopCulture® Reagent
| | • | 25 ml | Ni-NTA His•Bind Resin
| | • | 125 ml | 4X Ni-NTA Bind Buffer
| | • | 2 x 125 ml | 4X Ni-NTA Wash Buffer
| | • | 50 ml | 4X Ni-NTA Elute Buffer
| | • | 1 | 2-ml 96-well Filter Plate
| | • | 1 | 1-ml 96-well Collection Plate
with Sealer | | • | 300 KU | rLysozyme™ Solution
(Note: 1 KU = 1000 units) | | • | 1 ml | rLysozyme Dilution Buffer
| | • | 10,000 U | Benzonase® Nuclease,
Purity > 90% |
|
