Key information: Janelia Fluor® 570, NHS ester is an orange photosensitizer for proximity labeling experiments; supplied with an NHS ester reactive for the labeling of primary amines.
Suitable for live cell applications. Upon absorption of light energy, photosensitizers in the triplet excited state interact with molecular oxygen resulting in the formation of singlet oxygen and other ROS that can be leveraged to manipulate or interrogate biological systems.
Used for: proximity labeling experiments.
Application: photosensitizer-dependent oxidation and capture by amine (POCA) platforms reporting intracellular interactomes for both proteins and lipids such as cholesterol. Also suitable for chromophore-assisted light inactivation (CALI) of proteins in live cells; targeted ablation of cells; DAB photopolymerization for contrast in light and electron microscopy (EM).
Properties and Photophysical Data: NHS ester can be converted to relevant substrate for use in self-labeling tag systems, e.g. Haloalkane and SNAP-tag®. Excitation and emission maxima (λ) are 570 nm and 593 nm, respectively; quantum yield φ = 0.63; extinction coefficient ε = 83,600 M-1cm-1; singlet oxygen quantum yield φSO = 0.088; lactone-zwitterion equilibrium constant (KL-Z) = 2.24. Janelia Fluor® 570, NHS ester is net neutral and cell permeable.
Protocol: for cell-based POCA: cells are dosed with JF570-NHS (100 nM, 10 min) followed by washout to remove excess reagent (2 x 20 min), addition of PA (10 mM) and irradiation (170,000 Lux, 5 min,15 W yellow LED). After cell lysis, click chemistry and SDS-PAGE are performed.
Please see the protocol for further information on protein/antibody labeling and conjugation.
We also offer custom conjugation antibody services.
SNAP-tag® is a registered trademark of New England BioLabs, Inc.
Janelia Fluor® is a registered trademark of Howard Hughes Medical Institute.
