Sulfidefluor 7-AM  (Synonyms: SF7-AM)

Sulfidefluor 7-AM is a stable hydrogen sulphide (H₂S) fluorescent probe^[1]. Sulfidefluor 7-AM is a click chemistry reagent, it contains an Azide group and can undergo copper-catalyzed azide-alkyne cycloaddition reaction (CuAAc) with molecules containing Alkyne groups. It can also undergo strain-promoted alkyne-azide cycloaddition (SPAAC) reactions with molecules containing DBCO or BCN groups.

Guidelines (The following is a recommended protocol. This protocol is provided as a guide only and should be modified to suit your specific needs).^[2]
1. Preparation of Dye Stock Solution
1) Weigh an appropriate amount of dried SF7-AM probe and dissolve it in anhydrous DMF to prepare a 5 mM SF7-AM stock solution.
2. Preparation of Dye Working Solution
1) Dilute a volume of the 5 mM SF7-AM stock solution to a 2.5 μM working solution in complete cell culture medium. Prepare the working solution immediately before use and mix thoroughly.
3 Specific Staining Procedure
1) Two days before the imaging experiment, passage cells into phenol red-free medium and seed onto pre-coated (if necessary) four-well Lab-Tek II glass chamber slides. Culture until the cells reach 70%-80% confluency at the time of the experiment.
2) 60 min before imaging, remove the medium from each well and add 500 μL of 2.5 μM SF7-AM working solution. Incubate the cells at 37°C, 5% CO₂ for 30 min.
3) 30 min before imaging, remove the dye-containing medium and replace with fresh medium. For the positive control wells, add an appropriate amount of Na₂S stock solution to a final concentration of 100 μM. For the negative control wells, add an equal amount of water and continue incubating at 37°C, 5% CO₂ for 30 min.
4) Place the cells on the microscope stage and image the SF7-AM using an LSM 710 laser scanning confocal microscope with a 40x objective and a 488 nm Ar laser. Quantitative analysis was performed using Zen 2010 software, using uniform upper and lower thresholds to evaluate the mean pixel intensity of each image.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

685.55

C₃₁H₂₃N₇O₁₂

1416872-50-8

Solid

White to light brown

[N-]=[N+]=NC1=CC=C2C(OC3=CC(N=[N+]=[N-])=CC=C3C24OC(C5=CC(C(N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)=O)=CC=C54)=O)=C1

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Cheng Z, et al. Restoration of Hydrogen Sulfide Production in Diabetic Mice Improves Reparative Function of Bone Marrow Cells. Circulation. 2016;134(19):1467-1483.  [Content Brief]

  [2]. Lin VS, et al. Azide-based fluorescent probes: imaging hydrogen sulfide in living systems. Methods Enzymol. 2015;554:63-80.  [Content Brief]