HIS Lite(TM) iFluor(TM) 647 Tris NTA Chelator

Correction Factor (260 nm) 0.03. Correction Factor (280 nm) 0.03. Correction Factor (656 nm) 0.0793. Extinction coefficient (cm^-1 M^-1): 250000 (aqueous buffer (pH 7.2)). Excitation (nm): 656. Emission (nm): 670. Quantum yield 0.25 (aqueous buffer (pH 7.2)). iFluor(R) 647 Tris-NTA Chelator is used as a sensitive red fluorescent probe for detecting polyhistidine-labeled proteins in combination with the addition of a heavy metal ion such as Ni2+ in cells, solution and solid surfaces. Fluorescent tris-NTA compounds provide an efficient method for site-specific and stable noncovalent fluorescence labeling of polyhistidine-tagged proteins. In contrast to the transient binding of conventional mono-NTA, the multivalent interaction of tris-NTA conjugated fluorophores form a much more stable complex with polyhistidine-tagged proteins. The high selectivity of tris-NTA compounds toward cumulated histidines enables the selective labeling of proteins in cell lysates and on the surface of live cells. Fluorescent tris-NTA conjugates can be applied for the analysis of a ternary protein complex in solution and on surfaces. In combination with OG488-tris-NTA compound iFluor(R) 647 Tris-NTA can be used for multicolor analysis of polyhistidine-tagged proteins. AAT Bioquest also offers HIS Lite(TM) iFluor(TM) 647 Tris NTA-Ni Complex that can be directly used for the specific and highly sensitive detection of His-tagged fusion proteins without the addition of a heavy metal ion. The transition metal ions (e.g., Ni ion)-mediated complexation of polyhistidine-labeled proteins with fluorescent tris-NTA conjugates provides a sensitive reporter for detecting and monitoring protein-protein interactions in real time.