302104-25 302104-100
No Wash Fixable Viability- 488
BioMagis
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DESCRIPTION
No Wash Fixable Viability – 488 is a probe for binding phosphatidylserine (PS) exposed on the outer leaflet of the cell membrane on the dead cells in calcium independent manner. It can be used to quickly detect the dead cells with flow cytometry or fluorescent microscope. No cell wash needed. This probe can be used for multicolor staining with conjugated antibodies or dead cell indicators in the buffer without calcium. Also, this probe is fixable.
DETAILS
- Contains: One vial of Calcium Free Apoptotic-Dead Cell Tag – 488 One vial of DMSO One vial of dilution buffer
- Application: FC
- Fluorophore: A488
- 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. Gently add blocking buffer (PBS plus 5% FCS or BSA) for 15 minutes at room temperature. 3. Decant the blocking buffer. 4. Take the appropriate volume of reconstitute Calcium free Apo-dead Cell Tag solution. Recommend add 5 µL of reagent in 100 µL of staining buffer to 1 million cells for 15 minutes at room temperature in dark. Notes: The optimal concentration of Calcium Free Apoptotic-Dead cell tag may be different for different type of cells. 5. Gently wash the cells once. 6. Mount the cells with antifade mounting medium for imaging. Note: Cells can be fixed after staining.
- Excitationemission: Ex 488 / Em 520 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 can be stored at -20oC for six months. Once diluted to dilution buffer can be stored at 4-8oC for three months.
- Reconstitution and Usage: Place the vial at room temperature for a couple of minutes follow with a quick spin. • For 302104-100 / 100 Tests Reconstitute with 11 μl of DMSO, solution (pre- , 50x dilute to dilution buffer (1 μL to 50 μL dilution buffer). Recommend 5 μl per 100 μl of any staining buffer for 1 million cells. • For 302104-25 / 25 Tests Reconstitute with 130 μL of dilution buffer. Recommend 5 μl per 100 μl of any staining buffer for 1 million cells. • For 302104-20 / 20 Tests ICC staining Reconstitute with 110 μL of dilution buffer. Recommend 5 μl per 100 μl of any staining buffer for 1 million cells.