The SNAP-Capture Magnetic Beads are used to selectively immobilize and magnetically separate a SNAP-tag® fusion protein from solution using magnetic agarose beads. These beads show a low non-specific absorption of proteins from a complex lysate, making them suitable for pull down applications. The SNAP-Capture Magnetic Beads are prepared by the coupling of SNAP-tag substrate benzylguanine with highly stable 75-150 µm superparamagnetic particles. Two ml of SNAP-Capture Magnetic Beads is sufficient to perform 25 immobilization assays using 80 µl of bead suspension per assay.
The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule or modified surface to a protein of interest. The SNAP-tag is a protein based on mammalian O6-alkylguanine-DNA alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzyl purines and benzyl pyrimidines. In the immobilization reaction, the SNAP-tag is covalently attached to the substituted benzyl group of the SNAP-Capture Magnetic Beads.
There are two steps to using this system: sub-cloning and expression of the protein of interest as a SNAP-tag fusion, and capture and immobilization of the fusion protein using SNAP-Capture Magnetic Beads.
Materials Required but not Supplied
•Protein sample containing the protein to immobilize expressed as a SNAP-tag fusion
•Magnetic particle separator
•Buffer for immobilization and washing
Binding Capacity
SNAP-Capture Magnetic Beads (80 µl) were washed, incubated with 200 µl of 1 mg/ml SNAP-tag CBD (Chitin Binding Domain) for 1 hour at room-temperature, then rewashed as described in these instructions. Binding capacity was determined to be 3 mg/ml bead suspension.
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