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TAMRA, carboxyl tetramethy lrhodamine, is a luciferin derivative of rhodamine, which is widely used in the preparation of biological conjugated fluorophore, especially for fluorescent antibodies and avidin derivatives used in immunochemistry. TAMRA is one of the few fluorescent dye molecules that can be used to label proteins, so it is widely used as the marker of peptides and proteins.
Characteristics of TAMRA
TAMRA retains the typical chemical structure of rhodamine and contains three benzene rings. The balance between spironolactone and ring opening forms controls the conjugation of the ring. In nonpolar solvents, TAMRA dye exists in the form of spironolactone. Due to the interruption of heteroanthracene nucleus conjugation, its molar extinction coefficient (fluorescence quantum yield and lifetime) is very low. Lactone forms undergo charge separation in polar solution to form zwitterions. The open spiro ring can further form cations after protonation. Due to the ring opening process of chromophore conjugation, their absorption and fluorescence are greatly enhanced. In the ring opening form, rhodamine dyes exist in the form of ammonium ions and have good biocompatibility, which promotes the structural modification and modification of rhodamine dyes and has attracted extensive interest of researchers.
Application of TAMRA
Among the TAMRA series, 5-TAMRA and 6- TAMRA are most widely used. 5-TAMRA is dominant in polypeptide and protein labeling, while 6-TAMRA is often used in nucleotide labeling and nucleic acid sequencing. TAMRA monomer has gradually become the preferred reagent for labeling peptides and nucleotides because of its better resolution in HPLC (high performance liquid chromatography) purification, which is often used in the preparation of conjugates At the same time, 5 (6) - carboxytetramethylrhodamine (TAMRA, TMR) can be used as a fluorescent probe to study the molecular distribution of cell osmotic peptide chemical books (CPPS) and as a fluorescent component for two-dimensional fluorescence intensity distribution analysis (2d-fida), cold light resonance energy conversion (LRET) and fluorescence resonance energy conversion (FRET).
- 5-TAMRA can produce bright, pH insensitive orange fluorescence (excitation and emission extreme values are 546 / 579), and has good photostability. 5-TAMRA is widely used to prepare biological fluorescent groups, including peptides, proteins, nucleic acids, nucleotides, fluorescent antibodies, especially antibody fluorescent groups and avidin derivatives in immunohistochemistry. In addition, 5-TAMRA is widely used as a receptor for fam fluorescent groups in FRET (fluorescence resonance energy transfer) research.
- Under the name TAMRA, the carboxylic acid of 6-TAMRA has also achieved prominence as a dye for oligonucleotide labeling and automated DNA sequencing applications. 6-TAMRA is mainly used for automatic DNA sequencing and can react with primary or secondary amine groups in proteins, nucleic acids and other biomolecules.