What is the excitation and emission spectra of propidium iodide?
Propidium Iodide (PI) binds nucleic acids by intercalation in the bases. When bound to nucleic acid it has an excitation maxima ~535 nm and an emission maxima ~615 nm. On a flow cytometer PI is typically excited by 488 or 561 nm and can be detected in a 610/20 bandpass.
Is 7-AAD toxic to cells?
7-AAD is a membrane impermeant dye that is generally excluded from viable cells.
How do you use 7-AAD stain?
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.
Does propidium iodide stain apoptotic cells?
Propidium iodide is a membrane-impermeable fluorescent DNA stain. It is used to stain apoptotic cells for flow cytometry, in situ hybridization, and immunohistochemistry applications Belloc et al (1994). It can also be used to differentiate between apoptotic and necrotic cell death.
Can I use 7-AAD instead of propidium iodide?
7-AAD can be used in place of PI (propidium iodide). This solution can be used in combination with PE (phycoerythrin) and FITC (fluorescein isothiocyanate) conjugated antibodies in 2-color analysis. The advantage of 7-AAD over PI is that there is minimal spectral overlap between these emissions.
What is 7-AAD used for?
7-AAD (7-amino-actinomycin D) is a ready-to-use solution for the exclusion of nonviable cells in flow cytometric analysis. 7-AAD can be used in place of PI (propidium iodide). This solution can be used in combination with PE (phycoerythrin) and FITC (fluorescein isothiocyanate) conjugated antibodies in 2-color analysis.
What can be used in place of Pi (propidium iodide)?
7-AAD can be used in place of PI (propidium iodide). This solution can be used in combination with PE (phycoerythrin) and FITC (fluorescein isothiocyanate) conjugated antibodies in 2-color analysis.
How do you use 7 AAD dye?
Discrimination between viable and non-viable cells can be carried out with the use of the 7-AAD Viability Dye (Cat. No. 00-6993-50) or Propidium Iodide (PI) Staining Solution (Cat. No. 00-6990-50). The 7-AAD or PI will mark the non-viable cells by binding to the nuclei of those cells.
