Calcium Fluorescent Dyes Product Description Cell membrane potential fluorescent probe Calcium ion fluorescent probe FLUO-3AM application details First, the physical and chemical properties of Fluo-3AM Color and shape: orange red liquid purity: greater than 95% (high performance liquid phase) Carpet Binding Tape,Carpet Edge Binding Tape,Self Adhesive Carpet Binding Tape,Self Adhesive Carpet Binding Tape Kunshan Jieyudeng Intelligent Technology Co., Ltd. , https://www.jerrytape.com
DiBAC4(3) is a lipophilic anionic fluorescent dye sensitive to cell membrane potential. DIBAC4(3) itself has no fluorescence. When it enters the cell and binds to the protein in the cytoplasm, it emits fluorescence. DIBAC4(3) enters the cell and is intracellular. An increase in fluorescence intensity, that is, an increase in membrane potential indicates cell depolarization; conversely, a decrease in intracellular fluorescence intensity, that is, a decrease in membrane potential indicates hyperpolarization of cells. DiBAC4(3) is the most commonly used mitochondrial membrane potential detection fluorescent probe.
Solubility: soluble in DMSO
Absorption wavelength / emission wavelength: 506nm / 526nm (low / high calcium)
Dissipation coefficient: 86000/Mcm (506nm)
Storage and use: Powdered FLUO-3AM should be stored at -20 degrees or higher. It is recommended to use DMSO as a solvent. Both FLUO-3AM and DMSO must be allowed to stand at room temperature prior to dissolution. The dissolution process can be slow. Long-term storage of FLUO-3AM dissolved in anhydrous DMSO should be tightly sealed and stored at -20 °C. Before use, leave the preservation solution at room temperature before opening. The AM probe can be prevented from hydrolyzing during long-term storage.
Second, the working principle of Fluo-3AM FLUO-3 is widely used in long-wavelength fluorescent calcium indicator. It is absorbed at 506 NM and can be excited by the 488 nm excitation light of an argon ion laser for easy detection. FLUO-3 produced no fluorescence in the absence of binding to calcium. However, after binding to calcium, the fluorescence (526 nm) is enhanced by at least 40 fold. Fluo-3 AM is a fluorescent dye that penetrates cell membranes. The fluorescence of Fluo-3 AM is very weak and its fluorescence does not increase with increasing calcium ion concentration. Upon entering the cell, Fluo-3 AM can be cleaved by intracellular esterase to form Fluo-3, which is retained in the cell. Fluo-3 binds to calcium ions and combines with calcium ions to produce strong fluorescence with a maximum excitation wavelength of 506 nm and a maximum emission wavelength of 526 nm. The excitation wavelength recommended for actual detection is about 488 nm, and the emission wavelength is 525-530 nm.
Third, the application of Fluo-3AM
1. Process Overview For intracellular calcium ion detection, the usual concentration of Fluo-3 AM is 5 μM. Fluorescence probe loading is usually carried out by incubating with a suitable solution containing 5 μM of Fluo-3 AM and cells at 37 ° C for 30-60 minutes, followed by appropriate washing. After washing, consider appropriate incubation for another 20-30 minutes to ensure Fluo-3 AM. Complete conversion to Fluo-3 in cells. Detection is then performed on a flow cytometer.
2, Fluo-3AM use liquid configuration
100 ug of Fluo-3AM with a molecular weight of 1219.9. According to the actual molar concentration required, anhydrous DMSO dissolved. It is generally configured as a 1 to 5 mM DMSO stock solution. If it is insoluble, it can be solubilized with 20% F127 in DMSO.
Fourth, note:
1. Fluorescent dyes have quenching problems and should be kept away from light as much as possible;
2. Pay attention to personal protection during the experiment, wear gloves and masks;