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Dye to protein conjugation is a chemical strategy to form a stable covalent link between a dye and a protein, which is not destructible under most biological conditions. Due to more flexibility in choosing the labeling sites as well as enhanced fluorescence compared to proteins' intrinsic fluorescence, small organic dyes have been extensively used in protein labeling.
The biotin-streptavidin complex is one of the most popular tagging systems for the conjugation of biomolecules, such as proteins, lipids and nucleic acids, as well as that of synthetic molecules, such as fluorescent labels. It has found strong success in the area of sample preparation as a core part of many purification systems and plays a critical role in many detection systems for instruments such as microscopy and flow cytometry.
Buccutite™ crosslinking technology provides an efficient method to conjugate proteins with another macromolecule such as an antibody or an enzyme. More robust and with higher yield than the commonly-used SMCC, Buccutite™ crosslinking technology utilizes two exclusive linkers with unique properties. In general terms, these proprietary crosslinkers work as two halves of a single homobifunctional crosslinker. Each has an amine-reactive group on one end which specifically targets primary amines on the desired protein. The other end contains our proprietary Buccutite™-reactive group, which has a high degree of affinity for only binding its respective Buccutite™ MTA/FOL crosslinker counterpart. When each is present, the two conjugates covalently link together at their Buccutite™-reactive site to form a protein-protein conjugate.
Dye-labeled peptides and oligonucleotides are important tools in biochemical and cellular studies. Fluorescent peptides and oligonucleotides have been extensively used in all major types of fluorescence imaging including fluorescence resonance energy transfer (FRET). These labeled biomolecules are widely used for diagnosing infectious diseases based on the molecular beacon and other technologies. FRET peptides and oligonucleotides have been also used for cell analysis via fluorescence-associated cell sorting (FACS) either in vivo or in vitro for research and diagnostic purposes.
The most important characteristics of dye-labeled peptides and oligonucleotides are high sensitivity and non-radioactive detection. Using suitable quenchers, either classic or modern (such as the Tide Quencher™ series in the image above) can assist researchers with superior non-fluorescent imaging.
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