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DiR

CAS No. : 100068-60-8

MCE 站：DiR

产品活性：DiR 是一种长链碳菁染料。碳菁染料广泛用作亲脂性示踪剂，用于标记细胞、细胞器、脂质体、病毒和脂蛋白。

研究领域：Others

作用靶点：Fluorescent Dye

In Vitro: Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses ａnd lipoproteins. Long-chain carbocyanines which include DiO (DiOC18(3)), DiI (DiIC18(3)), DiD (DiIC18(5)) ａnd DiR, ａnd dialkyl aminostyryl dye DiA (4-Di-16-ASP) are used for labeling membranes ａnd other hydrophobic structures. DiIC16(3) has shorter alkyl substituents (C16) than DiI (C18). They have extremely high extinction coefficients, environmental dependent fluorescence ａnd short excited-state lifetimes in lipid environments. They are oils at room temperature ａnd weakly fluorescent in water but highly fluorescent ａnd quite photostable when incorporated into membranes or bound to lipophilic biomolecules. These optical characteristics make them ideal for staining the cytoplasmic membranes of cells. Once applied to cells, these dyes diffuse laterally within the plasma membrane, resulting in staining of the entire cell.
DiO, DiI, DiD ａnd DiR exhibit distinct green, orange, red ａnd infrared fluorescence, respectively thus facilitating multicolor imaging ａnd flow cytometric analysis of live cells . DiO ａnd DiI can be used with standard FITC ａnd TRITC filters respectively. Among them DiI ａnd its analogs are most frequently used since they usually exhibit very low cell toxicity. In addition, DiI is widely used for determining lipoproteins such as LDL ａnd HDL. The lipophilic aminostyryl dye DiA is also often used for neuronal tracing.
General Protocol
1. Preparing Stain Solutions of Di
a. Prepare DMF, DMSO or ethanol stock solutions: The stock solutions should be prepared in dimethyl formamide (DMF), dimethylsulfoxide (DMSO, or ethanol DMSO at 1-5 mM. DMF is preferable to ethanol as a solvent for Di. The stock solution should be used promptly. Any unused solution need to be aliquoted ａnd refrozen at least -20℃. Avoid repeated freeze/thaw cycle. The solution can be stored for 6 months.
b. Prepare working solutions: Dilute the stock solutions into a suitable buffer such as serum-free culture medium, HBSS or PBS to make 1 to 5 μM working solutions. We do not recommend storing the aqueous solution for more than one day.
Note: The final concentration of the working solution should be empirically determined for different cell types and/or experimental conditions.
2. Suspension cells
a. Centrifuge at 1000 g at 4℃ for 3-5 minutes ａnd then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL.
b. Add 1 mL of Di working solution, ａnd then incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4℃ for 3-4 minutes ａnd then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
3. 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 5-30 minutes.
d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

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