Updated: September 25, 2019

For research use onlyĂ
Table 1. Product information
Catalog no.

Product nameGC902

CopperGREEN™

AmountStorage upon receipt

Stability

50 nmol≤−20°C, keep desiccated1 year (when unopened×5and protected from light.

and stored as described.)1.
CopperGREEN is a fluorescent probe that specifically detects CuI ion. Copper is one of the trace metals
required by living organisms, and is an essential cofactor for enzyme activities of cytochrome c oxidase,superoxide dismutase, and tyrosinase. On the other hand, abnormal homeostasis of copper leads to severe
disorders such as Menkes syndrome and Wilson’s disease, or may be related to Alzheimer’s disease andcancers. Among two redox states of copper, CuI and CuII, CuI is dominant in the intracellular reducing
environment. Cell permeable CopperGREEN which has almost no fluorescence, fluoresces upon reaction
with the intracellular CuI. Therefore it is suitable for live cell imaging of CuI.

Figure 1. (left) Fluorescence emission spectra before and after the reaction to CuI ions. The graphshows spectra of 5 μM CopperGREEN in 50 mM Hepes, 2 mM glutathione (pH7.2). Fluorescenceincreases >100-fold upon reaction with CuI. (right) Metal ion selectivity of CopperGREEN in 50 mM
Hepes buffer (pH 7.2). Significant increase of fluorescence is observed only in the presence of CuI ions.
■ Storage
This product is shipped at room temperature. Upon receipt, store the product desiccated and protected fromlight at ≤ −20°C. Storing as a solution is not recommended.

2.
■ Materials required but not provided
・Dimethyl sulfoxide (DMSO)
・Observation buffer (1×PBS pH 7.4, Hank’s Balanced Salt Solution (HBSS), etc.). It should be a solution
without phenol red.

■ Preparation of reagent
1. CopperGREEN is a colorless solid. Before opening the vial, spin down the solid to the bottom by amicrocentrifuge.
2. Warm the vial to the room temperature and add 50 μl of DMSO to one vial to make 1 mM stock solution.
Dissolve the crystal completely by careful pipetting. Solution will be also colorless liquid. Use neutral buffer
solution for the dilution of the stock solution. CopperGREEN could be oxidized in acidic solutions.
■ Example cell imaging procedure
3. Prepare HeLa cells seeded on a glass-bottom dish and cultured for >12 hours at 37oC, 5% CO2. Add 100
μM CuCl2 to the cell culture medium (ex. DMEM + 8% FBS + P.S.) and culture for >12 hours. Then gently
rinse cells 2 times with PBS containing 200 μM EDTA to remove extracellular Cu2+.
4. Dilute the 1 mM CopperGREEN solution with the culture medium to 5 µM (Cell staining solution).
5. Replace the cell culture medium with the cell staining solution and incubate at 37oC, 5% CO2 for 3 hours.
6.
µM°C.

7. After the staining, rinse 2 times with the observation buffer.
8. Observe the cells with a fluorescent microscope.
※■ Fluorescence observation
CopperGREEN reacted with CuI is exited with lights around 440—490 nm wavelength, and emits
fluorescence with a peak at 510 nm. Use 488 nm laser or blue excitation filter cubes for GFP or FITC for
fluorescent microscopes
Table 2. Related Products
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