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Sigma HIS-Select* iLAP* Biomek® FX
Third-Party Application (system support)
Sigma's HIS-Select* iLAP* Plates for Integrated Lysis and Purification of Recombinant Histidine-tagged Proteins from Bacteria on the Biomek® FX
Sigma-Aldrich Corp.
Biotechnology Division
P.O. Box 14508, Saint Louis, MO 63178
www.sigmaaldrich.com
The Biomek® FX files provided along with this document were designed
to run Sigma’s HIS-Select* iLAP* HC Nickel Coated plates for the
integrated lysis and purification of recombinant histidine-tagged proteins
from bacteria. The following describes the hardware requirements necessary
to utilize these
methods, along with instructions on how to import the files. Before running
the automated method, please review the Automation Protocol for HIS-Select
iLAP plates on the Biomek FX located at www.sigmaaldrich.com
The HIS-Select iLAP (integrated Lysis and Affinity Purification) Plate
combines the cell lysis and purification steps into one. These plates
are coated with a proprietary high-density nickel chelate matrix. This
matrix provides high-capacity affinity binding of recombinant fusion proteins
with histidine-containing tags. An additional coating contains all of
the reagents and chemicals necessary for the lysis of bacterial cells
for the purification of recombinant proteins. In addition, it includes
protease inhibitors to help prevent the proteolytic breakdown of proteins.
These two coatings allow bacterial culture to be added directly to the
plate and simultaneously lysis cells and capture target protein. The iLAP
plates may be used to confirm expression of the his-tagged target protein,
protein-protein interaction assays, and to rapidly screen multiple constructs.
The resulting purified protein is compatible with protein assays, SDS-PAGE,
Western blotting, and MALDI.
* All trademarks are the property of their respective owners. Where applicable,
the PCR process is covered by patents owned by Roche Molecular Systems,
Inc., and F. Hoffman-LaRoche, Ltd.
