RhFNMB 

RhFNMB is a dualchannel/localization single-molecule fluorescence probe for ATP and HOCl, with independent fluorescence responses in the light red channel with ATP (λex = 520 nm, λem = 586 nm) and deep red channel with HOCl (λex = 620 nm, λem = 688 nm)^[1].

General Protocol
1. Preparation of RhFNMB working solution
1.1 Prepare a 10 mM stock solution
1.2 Prepare a 5-10 μM RhFNMB working solution
Note: Please adjust the concentration of RhFNMB working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells (6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 min and then discard the supernatant. Wash twice with PBS, 5 min each time.The cell density is 1×10⁶/mL.
b. Add 1 mL of working solution, and then incubate at room temperature for 1 h.
c.Centrifuge at 400 g at 4°C for 3-4 min and then discard the supernatant.
d. Wash twice with PBS, 5 min each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 1 h.
d. Wash twice with medium, 5 min each time. Observation by fluorescence microscopy or flow cytometry.
Precautions
1. Please adjust the concentration of RhFNMB working solution according to the actual situation.
2. This product is only for R&D use, not for drug, household, or others.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

882.17

C₅₁H₆₃N₉O₃S

CCN(C1=CC=C2C(OC3=C(C24C5=C(C(N4CCNCCNCCNC(N6C7=C(SC8=C6C=CC(N(C)C)=C8)C=C(C=C7)N(C)C)=O)=O)C=CC=C5)C=CC(N(CC)CC)=C3)=C1)CC

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Qu W, et al. Dual-Channel/Localization Single-Molecule Fluorescence Probe for Monitoring ATP and HOCl in Early Diagnosis and Therapy of Rheumatoid Arthritis. Anal Chem. 2024 Apr 9;96(14):5428-5436.  [Content Brief]