What does 7-AAD stain for?
7-aminoactinomycin D (7-AAD) is a fluorescent intercalator that undergoes a spectral shift upon association with DNA. 7-AAD/DNA complexes can be excited by the 488 nm laser and has an emission maxima of 647 nm, making this nucleic acid stain useful for multicolor fluorescence microscopy and flow cytometry.
How do you use 7-AAD in flow cytometry?
7-AAD stock buffer: Dissolve 1 mg of 7- AAD powder by adding 50 microliters of absolute methanol directly to the vial. Mix well and add 950 microliters of 1 X PBS with Ca2+ and Mg2+ to achieve a concentration of 1 mg/ml. Store solution tightly closed and protected from light at 4°C.
Can 7-AAD be fixed?
7-AAD is widely used for live/dead cell discrimination. It also can be used in fixed and permeabilized cells for cell cycle profiling by DNA content analysis using flow cytometry. 7-AAD intercalates selectively at GC-rich regions of DNA, making it dye useful for chromosome banding studies.
Is 7-AAD toxic?
I didn’t observe any toxicity due to 7AAD in the samples and results are just fine everytime. I will recommend using them together to avoid the over complications in protocol. 7AAD binds well in Annexin binding buffer also. You can wash the dyes off by PBS once incubation is completed.
What color is 7-AAD in?
Red
7-Aminoactinomycin D
| Names | |
|---|---|
| Chemical formula | C62H87N13O16 |
| Molar mass | 1270.43 g/mol |
| Appearance | Red to dark purple powder |
| Hazards |
What channel is 7-AAD?
The fluorescence emission maximum for 7-AAD is at 647 nm. When excited at 488 nm, 7-AAD is detected in the red fluorescence channel commonly used for R-phycoerythrin (PE)-Cy®5 tandem dye detection, with minimal spectral overlap into the yellow fluorescence channel commonly used for PE detection.
What is PerCP in flow cytometry?
Peridinin Chlorophyll Protein Complex (PerCP)-conjugated antibodies are convenient tools for use in flow cytometry experiments. PerCP is a water soluble carotenoid pigment found in photosynthetic dinoflagellates.
What color is PerCP?
PerCP-Cyanine® 5.5 (PerCP-Cy5. 5) is a red-emitting tandem fluorophore that combines PerCP and Cy5. 5. The donor molecule, PerCP can be excited by the 488-nm blue laser and and transfers energy to the acceptor molecule, Cy5.
What channel is DAPI in flow cytometry?
With an excitation maximum of 358 nm DNA-bound DAPI can be excited by an ultraviolet (UV) laser or a violet (405 nm) laser and detected within the blue fluorescence channel, e.g., channel V1 on the MACSQuant® Analyzer 10. Exclusion of dead cells from flow cytometric analysis.
What is PE and APC in flow cytometry?
Fluorophores for flow cytometry. * PE is the same as R-phycoerythrin. APC, allophycocyanin; FITC, fluorescein isothiocyanate; PE, phycoerythrin; PerCP, peridinin chlorophyll protein.