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Updated: Dec 24, 2025
Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
Published on: May 17, 2010
A Ni-NTA-based red fluorescence probe for protein labelling in live cells
Ailun Chao1, Nan Jiang, Ya Yang
1Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong, P. R. China. hsun@hku.hk.
Abstract:
The great success of a His6-Ni-nitrilotriaceate (Ni-NTA) system in protein purification has inspired scientists to develop novel Ni-NTA based fluoresent probes for imaging of proteins in live cells. Despite a great deal of effort being made, only very few such probes can cross the cell membrane to label intracellular proteins. Herein, a new red fluorescence probe (Ni-NTA-AB) consisting of a Ni-NTA moiety, BODIPY and an arylazide was designed and synthesized to selectively label His6-tagged proteins. Guided by the Ni-NTA, the probe selectively binds to proteins with a genetically fused His6-tag exemplified by a DNA repair protein, xerodermapigmentosum group A (XPA122), leading to ca. 26-fold fluorescence enhancement. Ni-NTA-AB could enter into live bacterial cells to label intracellular His6-tagged proteins with the assistance of Tween 80 and also to quantify His6-tagged proteins in SDS-PAGE upon photoactivation of arylazide with a wide linear range (25-1000 ng). This study may provide a new horizon for the multiple-channel tracking and labelling of His6-tagged proteins.
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