Cells often respond to environmental signals by synthesizing new proteins, forming new protein complexes, or post-translationally modifying existing proteins. The ProteoTuner Immunoprecipitation Kit gives you the ability to selectively distinguish these novel events from the existing protein pool, and to better understand cellular responses to external stimuli.
The kit allows you to choose when, and for how long, to stabilize protein complexes before you immunoprecipitate them, so it can be used to increase the signal-to-noise ratio for many types of IP studies, including signal transduction studies.
How the ProteoTuner Immunoprecipitation Kit Works
Clontech’s ProteoTuner Immunoprecipitation Kit is a novel application of ProteoTuner protein stabilization technology, which enables you to clone a protein of interest in frame with a 12 kD destabilization domain (the DD tag). Once your ProteoTuner vector (sold separately) is transfected into cells, your protein of interest is constitutively expressed—but also continually targeted to the proteasome and degraded. However, adding the membrane-permeable small molecule Shield1 to the culture medium protects the DD fusion protein from being degraded, and allows it to accumulate in a dose-dependent manner. The ProteoTuner Immunoprecipitation Kit provides all the materials you need to IP the newly accumulated protein and co-purify its interacting partners.
Magnetic Bead-Based IP Protocol
The kit contains an anti-DD monoclonal antibody (Clone 24A) that has been selected for its ability to immunoprecipitate very efficiently, lysis and wash buffers, and magnetic IgG beads. Simply mix your lysate with the antibody and pull down using these Mag Capture Beads and a magnetic separator such as the Magnetic Stand for Microcentrifuge Tubes (sold separately; Cat. No. 631964).