English
中文
CAS:100068-60-8
Storage:Powder:2-8℃，2 years;Insolvent(Mother Liquid):-20℃，6 months;-80℃，1 year
Purity:≥98%
Appearance:Solid
Cy7 DiC18 (DiR) is a lipophilic, near-infrared fluorescent cyanine dye. This dye is often used to label the plasma membrane of cells. The two 18-carbon chains of Cy7 DiC18 (DiR) are inserted into the cell membrane for specific, stable cell staining with virtually no intercellular dye transfer. Cy7 DiC18 (DiR) (near infrared fluorescence), used in combination with other cell membrane fluorescence dyes such as DiI (orange fluorescence), DiO (green fluorescence), and DiD (red fluorescence), provides an effective tool for multi-color imaging and flow cytometry. Cy7 DiC18 (DiR) can be fixed with paraformaldehyde after staining (no other reagents such as methanol can be used), but it is not recommended to undergo the process of penetration after dyeing. In addition, plasma membrane staining can also be performed well after fixed penetration (penetration with 0.1% TritonX-100 at room temperature).
How to use (for reference only)
Dyeing solution preparation
(1) Preparation of storage liquid: the storage liquid is prepared with anhydrous DMSO or anhydrous EtOH, the concentration of 1~5 mM. Note:
Unused storage liquid is recommended to be stored at -20℃ after packaging to avoid repeated freezing and thawing.
(2) Preparation of working liquid: Dilute the storage liquid with a suitable buffer (such as serum-free medium, HBSS or PBS) to prepare a working liquid with a concentration of 1 to 5 μM. Note:
The final concentration of working fluid is recommended to be optimized according to different cell lines and experimental systems. It is recommended to search for the optimal concentration from a range of 10 times the recommended concentration.
Suspension cell staining
(1) Adding appropriate volume of dye working fluid to re-suspension cells, making their density 1×106/mL.
(2) Incubate the cells at 37℃ for 2~20 min, and the optimal culture time was different for different cells. 20 min can be used as the initial incubation time, and then the system can be optimized to obtain uniform labeling effect.
(3) After incubation, centrifuge at 1000-1500 rpm for 5 min. Pour the supernatant and slowly add the growth medium preheated at 37℃ again to resuspend the cells.
(4) Repeat step (3) more than twice.
Adherent cell staining
(1) The adherent cells were cultured on a sterile cover slide.
(2) Remove the cover glass from the medium to absorb excess fluid, but keep the surface moist.
(3) Add 100 μL of dye working liquid to one corner of the cover glass, and gently shake the dye to evenly cover all cells.
(4) Cells were incubated at 37℃ for 2~20 min, and the optimal culture time was different for different cells. 20 min can be used as the initial incubation time, and then the system can be optimized to obtain uniform labeling effect.
(5) Blot the dye working solution, wash the slide with the culture solution for 2 to 3 times, cover all cells with the pre-warmed medium each time, incubate for 5 to 10 min, and then blot the medium. But keep the surface moist.
Result detection
The sample can be detected in the medium and can be analyzed by fluorescence microscopy imaging or flow cytometry.
Matters needing attention
When staining fixed cell or tissue samples with Cy7 DiC18 (DiR), 4% paraformaldehyde prepared in PBS is usually used for fixation, and the use of other inappropriate fixation fluids will result in a higher fluorescent background.
