WSP-1

WSP-1 is a selective and rapid-reaction H₂S specific fluorescent dye (Ex/Em=465/515 nm). WSP-1 reacts with H₂S with the releasing of fluorophore^[1]. In Vitro:WSP-1 have fluorescence properties, with excitation wavelength=465 nm, emission spectrum=515 nm^[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Fluorimetric Detection of Intracellular H₂S (Fluorimetric Microscope Assessment)^[1]:
1. Culture HASMCs up to about 90% confluence, and 24 h before the experiment, seed cells onto a 8-well culture slide at density of 30*10³ per well and culture for 24 hours.
2. Replace the culture medium, and incubate the cells for 30 min at 37°C in 400 μL per well WSP-1 working solution for 30 min.
3. Replace the WSP-1 working solution with 380 μL of buffer standard and add 20 μL of the tested H2S-donor compounds at the desired concentration.
4. When WSP-1 reacts with H₂S, it releases a fluorophore detect able at Ex/Em=465/515 nm. Remove the buffer standard, and wash with 400 μL of DPBS after 1 h incubation.
5. Remove the DPBS, and add 200 μL of Bouin solution per well for 10 min at room temperature to fix the cells. Remove the excess of Bouin solution and wash the cells twice with 400 μL of DPBS.
6. Acquire the fluorescence signal with a fluorescence microscope.