Zinquin ethyl ester

Zinquin ethyl ester is a cell-permeable and lipophilic fluorescent derivative of Zinquin (HY-D0982). Zinquin ethyl ester reacts with protein-bound Zn²⁺ in cells and forms fluorescent ternary adducts. Zinquin ethyl ester undergoes hydrolysis by intracellular esterases impeding its efflux across the plasma membrane (Ex/Em = 370/470 nm)^[1]. In Vitro:Guidelines (The following is our recommended protocol; this protocol serves as a guideline only and should be modified to suit your specific needs.)
1. Preparation of Stock and Working Solutions
a. Prepare a 10 mM Zinquin ethyl ester stock solution in anhydrous DMSO.
b. Dilute the stock solution using pre-warmed, serum-free cell culture medium or PBS to prepare a 25 μM Zinquin ethyl ester working solution.
Note: Please adjust the concentration of the Rhobo6 working solution according to your specific experimental requirements, and prepare it immediately before use.
2. Cell Staining
a. Culture cells on sterile coverslips.
b. Remove the coverslips from the culture medium and aspirate any excess medium.
c. Add the working solution, gently agitating to ensure it completely covers the cells, and incubate at room temperature for 30 minutes.
d. Aspirate the working solution, then wash 2–3 times with culture medium (5 minutes per wash). Observe using a fluorescence microscope or flow cytometer.
Note: If analysis via flow cytometry is required, the cells must be detached using trypsin digestion and resuspended prior to staining.
Zinquin ethyl ester (25 μM; 30 min) forms fluorescent ternary adducts with Zn-proteins in LLC-PK1, TE671, U87 mg, C6, CCRF-CEM, A549, and mouse primary brain cells, producing a characteristic emission maximum near 470 nm with cell-type specific fluorescence intensities^[1].
Zinquin ethyl ester (25 μM) reacts primarily with proteomic Zn²⁺ in LLC-PK1, TE671, U87 mg, C6, CCRF-CEM, A549, and mouse primary brain cells to form ternary Zinquin acid-Zn-protein adducts^[1].
Zinquin ethyl ester (1-20 μM; 15 min) forms fluorescent ternary adducts with the isolated Zn-proteome from TE671 cells^[1].