Analyzing protein function is a key goal for discovery-based cell biology research. Regulating protein level rapidly in the cell as a method of accurate functional analysis is possible using ProteoTuner technology. The protein's level can be measured either by using the DD Antibody or the ProteoTuner Quantitation System.
The ProteoTuner DD Monoclonal Antibody recognizes the DD tag at the N or C-terminal of the protein and can be used for Western Blot Analysis and Immunocytochemistry.
ProteoTuner Quantitation System
The ProteoTuner Quantitation System combines precise control of the level of a protein of interest with a direct means to quantitate the degree of stabilization of this protein, without the requirement for a specific antibody and a lengthy Western blot protocol.
Rapid, Precise Control of Gene Induction
The ProteoTuner system controls the amount of your protein of interest by acting directly on the post-translated protein. Based on a tag (DD) and ligand (Shield1) technology, it enables you to reversibly “tune” the amount of stabilized, DD-tagged protein present in the cell very precisely, by varying the amount of Shield1 added to the culture medium.
Simple, Sensitive Protein Quantitation
The ProteoTuner quantitation kit bears a small 6 kDa ProLabel tag that is also tagged to the protein of interest. Using an enzyme fragment complementation assay in which two nonfunctional fragments combine in vitro to form a complete, active enzyme that cleaves a chemiluminescent substrate. The resulting signal is directly proportional to the amount of protein in a sample containing a protein bearing the ProLabel tag.
A Complete System for Protein Control & Quantitation
The ProteoTuner Quantitation Vector combines the ProteoTuner and ProLabel tags in one vector. It includes a multiple cloning site (MCS) to clone in the gene encoding your protein of interest, flanked on its N-terminus by the DD coding sequence and on its C-terminus by the ProLabel coding sequence. Using the ProteoTuner Quantitation System is simple: first, insert your gene of interest (without a stop codon) into the vector’s multiple cloning site, and transfect the construct into your experimental cell line. Then when you are ready to begin your experiment, stabilize your fusion protein with Shield1. To confirm the degree of protein stabilization that you have achieved, lyse a portion of your cells, combine them with the ProLabel detection reagents, and measure the resulting chemiluminescence.