Calcium Free Apoptotic-Dead Cell Tag - 647

DESCRIPTION

Calcium Free Apoptotic-Dead Cell Tag – 647 is a probe specifically designed to bind to phosphatidylserine (PS) exposed on the outer leaflet of the cell membrane in apoptotic cells and dead cells, regardless of calcium levels. This probe is suitable for detecting apoptotic cells, dead cells, activated platelets, and extracellular vesicles using flow cytometry or fluorescence microscopy. Moreover, it can be used in multicolor staining alongside conjugated antibodies or dead cell indicators in calcium-free buffers. Importantly, the staining performed with the calcium-free apoptotic-dead cell tag is fixable, enabling the distinction of viable cells for subsequent intracellular staining.

DETAILS

Application: FC, ICC
Fluorophore: Atto 647
Formulation: Lyophilized
Application Notes: Protocol for flow cytometer 1. Add 5 µL of the diluted reagent to 1 million cells in 100 µL of any staining buffer. No cell wash needed. 2. Incubate at room temperature for 5 minutes in the dark. 3. No wash needed. Directly running on the cytometer. Note: Different cell types can have a wide degree of variability in staining based on cell size and degree of cell death. Titrate the amount of dye and/or number of cells per 100 µl for optimal performance. Note: Cells can be fixed after staining. Protocol for ICC 1. Gently remove the buffer from the adherent cells 2. Take the appropriate volume of reconstitute Calcium free Apo-dead Cell Tag-dye solution. Recommend add 15 µL of reagent in 100 µL of staining buffer (containing 5% FCS or BSA) for 15 minutes at room temperature in dark. Notes: The optimal concentration of Calcium Free Apoptotic-Dead Cell Tag-dye may be different for different type of cells. 3. Gently wash the cells once. 4. Mount the cells with antifade mounting medium for imaging. Note: Cells can be fixed after staining.
Excitationemission: Ex 646 / Em 664 nm
Verified Reactivity: Human, Mouse
Storage and Handling: • Store at -20°C upon receipt. Do not open vials until needed • DMSO reconstitute solution of Apoptotic-Dead Cell Tag-dye can be stored at -20°C for up to up to six months. • Once diluted to dilution buffer can be stored at 2-8°C for up to three months
Reconstitution and Usage: Place the vial at room temperature for a couple of minutes follow with a quick spin. Reconstitute with 130 µL of dilution buffer. Recommend 5 µl per 100 µl of any staining buffer for 1 million cells