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Protein Regulation with Rapid Kinetics-ProteoTuner Systems

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Rapidly regulate the quantity of your protein of interest, at the post translational level. The ProteoTuner systems use a powerful, unique technology to control your protein in living cells. ProteoTuner technology has been successfully used with cytosolic and membrane-bound proteins in a variety of cell types, organisms, and animal models.

 

One Vector/One Ligand System

  1. A 12 kDa destabilizing domain (DD) that, when fused to a protein of interest, destabilizes the protein by targeting it for proteasomal degradation. Plasmid, retroviral and lentiviral PTuner vectors are available that carry the DD domain for fusion at either the N- or C-terminal of your protein.
  2. A membrane-permeable small molecule (750 Da) ligand, Shield1, which protects the DD fusion protein from being degraded and allows it to accumulate in the cell. Stabilization of the DD-protein occurs in as little as 15–30 minutes.

We also offer the ProteoTuner Tag Kit, which provides you with a ProteoTuner tag sequence so you can quickly and easily convert any Living Colors fluorescent protein-C vector into a ProteoTuner vector. The Kit contains a ProteoTuner destabilization domain (DD) source vector plus a primer mix that lets you PCR-amplify the DD tag sequence. You can then use an In-Fusion Cloning Kit to insert the DD tag sequence upstream of the fluorescent protein coding sequence in any of our fluorescent protein-C vectors. The resulting vector will be able to express a protein of interest that is N-terminally-tagged with a fluorescent protein and subject to ligand-dependent degradation.


Characterization:

Features

  1. Rapid kinetics: Protein level changes in minutes allows accurate functional analysis
  2. Precise tuning: Precise control of protein level is key to analyzing biological activity
  3. Reversible control: ‘Protein on’ to ‘off’ and back makes multiple studies easier
  4. Gets rid of accumulated protein: On demand rapid degradation of accumulated protein is important for analysis of essential proteins

Applications

  • Protein function in pathways
  • Functional analysis of subunits of a protein complex
  • Functional analysis of essential proteins


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