Site-Directed Fluorescence Labeling, TAMRA-C6-AF (amber), CloverDirect

Cosmo Bio LTD

Catalog No: PRX-CLD02

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About this maker: This item is a product of ProteinExpress Co. Ltd (Chiba, Japan), an R&D company expert cell-free protein synthesis and large scale cellular protein expression. See all ProteinExpress products at the ProteinExpress dashboard page.

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PRX-CLD02 is a TAMRA-C6-AF-charged aminoacyl-tRNA that recognizes and suppresses the UAG (amber) codon. CloverDirect™ tRNA Reagents for Site-Directed Protein Functionalization are designed for investigators who wish to incorporate unnatural amino acids at genetically defined positions of proteins using cell-free coupled transcription/translation systems, such as PUREfrex^®, PURESYSTEM^® and The RYTS Kit.

The targeting of unnatural amino acids is genetically defined by selectively introducing UAG amber or CGGG four-base codons into translation templates. For template construction, we recommend expression plasmids from Protein Express (pROX-FL92.1: ProX-tag + His-tag) or BioComber (pURE1-4: no tags). The ProX tag is a special N-terminal peptide designed to promote incorporation of certain large unnatural amino acids requiring precise N-terminal positioning (codon 9) and a specific codon context (flanked by AAC and TCT). Codon 9 of the ProX tag can be varied between UAG, CGGG or TTT (as a control).

During cell-free translation, unnatural amino acid-charged aminoacyl-tRNAs recognize either UAG amber or CGGG codons. If a UAG codon is recognized by release factor 1 (one of three termination factors), protein synthesis will be terminated. If the first three nucleotides of CGGG are recognized as a triplet codon by Arg-tRNA, the reading frame shifts to +1 and a downstream stop codon will terminate protein synthesis. Therefore, full-length translation products will contain unnatural amino acids at 100% efficiency.

Product contents

• Unnatural aminoacyl-tRNA (One tube of unnatural aminoacyl-tRNA is sufficient for 300μL of in vitro translation reaction.)
• tRNA buffer

Storage and stability

• Always store this product in the dark.
• This product is stable at -70°C until the expiration date which is indicated on the outside of the package.
• Once unnatural aminoacyl-tRNA is dissolved in tRNA buffer, it can be stored at -70°C for 2 months.

Applications

• Fluorescent labeling
• Biotin labeling
• PEGylation
• Chemical crosslinking
• Photo-regulation of a variety of biological processes, including enzyme, receptor, and ion channel activity
• In addition, upon request custom services for protein expression with unnatural amino acids are available.