Filtration-based, 96-well format purification directly from transfected cultures of insect cells

The Insect RoboPop™ Ni-NTA His•Bind^® Purification Kit is designed for filtration-based, 96-well format purification of His•Tag® fusion proteins directly from transfected insect cell cultures. This kit includes Insect PopCulture^® Reagent for protein extraction from total cultures, Benzonase^® Nuclease for viscosity reduction, Ni-NTA His•Bind Resin and buffers, 2-ml 96-well Filter Plate and Collection Plate with Sealer. The kit is configured for robotic processing of transfected 10-ml suspension cultures and purifies up to 400 µg His•Tag fusion protein per culture based on binding capacity of the resin. Protein yields ranging from 60 to 140 µg per 10 ml culture were obtained with b-gal, Fluc, MAP kinase, and cdc2 kinase expressed in the pIEx™ transient protein expression system (Loomis 2002).

In practice, Insect PopCulture Reagent is added directly to the cell culture, followed by the addition of Benzonase Nuclease. The lysate is transferred to a tube of appropriate size or deep-well plate and Ni-NTA His•Bind resin is added. After mixing, the lysate-resin slurry is transferred to the Filter Plate, and washing and elution steps are carried out on a vacuum manifold. The Filter Plate is compatible with standard vacuum manifolds for either manual or automated processing, and the entire purification has been validated for robotic sample processing with the Packard-brand MultiPROBE^® II liquid handling workstation from PerkinElmer Life Sciences and with the Genesis^® Freedom™ workstation from Tecan. Also use the Insect RoboPop Ni-NTA His•Bind Purification Kit with centrifugation when a vacuum manifold is not available.


Filter Plate
The 2 ml 96-well Filter Plate that can be used with the filtration-based RoboPop Purification Kits is also available separately.