High-throughput, milligram-scale purification of His•Tag^® and GST•Tag™ fusion proteins

The RoboPop™ GST•Bind™ Purification Kit is designed for high-throughput (HT), filtration-based purification of soluble GST•Tag™ fusion proteins directly from E. coli cultures. Like the corresponding RoboPop GST•Mag™ Kit (Grabski 2002), this kit features PopCulture® Reagent, rLysozyme™ Solution, and Benzonase® Nuclease for centrifuge-free cell lysis and extract preparation in one step. This kit is designed for larger-scale cultures (up to 5 ml) and includes GST•Bind Resin plus a filtration-based processing protocol. Whereas the magnetic-based GST•Mag Kits purify up to 50 µg target protein per 1 ml culture, the filtration-based kits purify up to 0.8 mg GST•Tag fusion protein per well.

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 MultiPROBE® II liquid handling workstation from PerkinElmer Life Sciences and with the Genesis® Freedom™ workstation from Tecan. A 96-well Collection Plate (1-ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins.

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%