Fluo-3FF AM 

Fluo-3FF AM is a low affinity (Kd = 42 μM) fluorescent Ca²⁺ sensitive indicator (Abs/Em = 462 nm/526 nm). Fluo-3FF AM is Mg²⁺ insensitive and relatively photostable. Fluo-3FF AM is an analog of Fluo-3FF AM. Fluo-3FF AM is essentially non-fluorescent, but exhibits a strong fluorescence enhancement upon entry into cells and binding to calcium.^[1]

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
1. Preparation of Fluo-3FF AM Solution 1.1 Preparation of Storage Solution Prepare a 1 mM stock solution by diluting Fluo-3FF AM with a 0.025% (w/v) Pluronic F-127/dimethyl sulfoxide solution.
1.2 Preparation of Working Solution Prepare a 5 μM working solution of Fluo-3 AM using a balanced buffer solution.
Note: Please adjust the concentration of the Fluo-3FF AM working solution according to the actual situation, and prepare it as needed.
2. Cell Staining 2.1 Cultivate adherent cells on sterile coverslips.
2.2 Remove the cover glass from the culture medium and aspirate away the excess medium.
2.3 Add 100 μL of the dye working solution and gently shake to ensure complete coverage of the cells. Incubate for 60 minutes.
a) Remove the dye working solution and then wash it in PSS at 4 °C for 60 minutes.
b) Imaging/Detection: The excitation wavelength is approximately between 462 - 506 nm, and the emission signal is detected at approximately 526 nm.

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

1151.80

C₅₀H₄₆Cl₂F₂N₂O₂₃

348079-13-0

O=C(C)OCOC(CN(CC(OCOC(C)=O)=O)C1=C(OCCOC2=C(C(F)=CC=C2N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)F)C=C(C3=C4C=C(C(C=C4OC5=C3C=C(Cl)C(OCOC(C)=O)=C5)=O)Cl)C=C1)=O

Room temperature in continental US; may vary elsewhere.

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

• [1]. Gordienko DV, et al. Regulation of muscarinic cationic current in myocytes from guinea-pig ileum by intracellular Ca2+ release: a central role of inositol 1,4,5-trisphosphate receptors. Cell Calcium. 2004 Nov;36(5):367-86.  [Content Brief]

  [2]. Young RC, et al. Focal sarcoplasmic reticulum calcium stores and diffuse inositol 1,4,5-trisphosphate and ryanodine receptors in human myometrium. Cell Calcium. 1999 Jul-Aug;26(1-2):69-75.  [Content Brief]