HIS Lite(TM) Cy5 Bis NTA Chelator

Correction Factor (260 nm) 0.02. Correction Factor (280 nm) 0.03. Correction Factor (482 nm) 0.009. Correction Factor (565 nm) 0.09. Extinction coefficient (cm^-1 M^-1): 250000 (ethanol). Excitation (nm): 651. Emission (nm): 670. Quantum yield 0.27 (PBS), 0.4 (ethanol). Cy5 Bis NTA-Ni Chelator can be applied either to an SDS-PAGE gel or Western blot membrane for fluorescence imaging or used in living cells in combination with the addition of a heavy metal ion such as Ni2+. Detection with Cy5 Bis NTA-Ni complex requires less incubation time than for protein-antibody binding. No secondary reaction is required since Ni-NTA complex is directly conjugated to the Cy5 fluorophores. Cy5 demonstrates strong fluorescent signals at ~670 nm with a common red laser excitation (e.g., He-Ne laser or 647 nm red laser). Polyhistidine is one of the most popular affinity tags incorporated into recombinant proteins. It can be inserted either at the N- or C-terminus and expressed in a variety of hosts. Due to its small size, the polyhistidine tag serves as an elegant tool for both protein purification and detection. AAT Bioquest also offers HIS Lite(TM) Cy5 Bis 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.