It is possible to introduce non-natural amino acids at targeted positions【CloverDirect】.
By simply adding this product to a cell-free translation system, you can efficiently synthesize proteins with non-natural amino acids in a short time.
○Quantitative: Non-natural amino acids can be introduced only at the desired position. ○Rapid and simple: Pinpoint labeled proteins can be obtained purified in about 2 hours. ○Variety: Non-natural amino acids labeled with fluorescence, biotin, etc., can be introduced.
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basic information
CloverDirect is a reagent used to introduce non-natural amino acids at specified sites in proteins utilizing a cell-free translation system. Non-natural amino acids with side chains such as fluorescent groups, biotin, PEG chains, and crosslinking agents, as well as post-translationally modified amino acids, are available. By simply adding a gene that incorporates one of the stop codons, such as the amber codon or a four-base codon, along with this product to the cell-free translation system, proteins with non-natural amino acids can be efficiently synthesized in a short time (technology introduced from the Japan Advanced Institute of Science and Technology).
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Applications/Examples of results
Product Usage Examples 1) A protease inhibitor discovery method using fluorescence correlation spectroscopy with position-specific labeled protein substrates. Anal Biochem. 2009 15;390(2):121-5. 2) “Quenchbodies”: quench-based antibody probes that show antigen-dependent fluorescence. J Am Chem Soc. 2011 2;133(43):17386-94. 3) Dynamically varying interactions between heregulin and ErbB proteins detected by single-molecule analysis in living cells. Proc Natl Acad Sci USA. 2012 28;109(35):13984-9. etc.
Detailed information
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Case Study 2UAG: Insertion of UAG codon immediately after the start AUG ProX tag*: Addition of ProX tag (UAG) Fluorescent imager detection wavelengths (above figure) HiLyteFluor488: Excitation 488nm / Detection 520nm TAMRA: Excitation 532nm / Detection 580nm ATTO633: Excitation 635nm / Detection 670nm Western blot (below figure): Anti-His antibody (Biotin-tagged protein uses anti-biotin antibody)
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■Construction of protein expression plasmids and protein expression/purification ■Manufacturing and sales of non-natural amino acid incorporation protein synthesis reagents (CloverDirect series) and E. coli cell-free translation kits (RYTS Kit) ■Sales of affinity ligands and affinity resins for antibody purification ■Sales of anti-KIAA polyclonal antibodies