No Wash Fixable Viability- 647

DESCRIPTION

No Wash Fixable Viability– 647 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

Application: FC, ICC
Fluorophore: A647
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 No wash fixable viability-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 No wash fixable viability-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 650 / Em 665 nm
Verified Reactivity: Human, Mouse
Storage and Handling: Store at -20°C upon receipt. Do not open vials until needed. DMSO reconstitute solution of No wash fixable viability-dye can be stored at -20°C for 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. • For 302093-100 / 100 Tests Reconstitute with 11 µl of DMSO, mix well, follow by another quick spin. Take the appropriate volume of DMSO solution (pre-calculate the volume as desired tests plus died volume), 50x dilute to dilution buffer (1 µL to 50 µL dilution buffer). Recommend 5 µl per 100ul of any staining buffer for 1 million cells. • For 302093-25 / 25 Tests Reconstitute with 130 µL of dilution buffer. Mix well, follow by another quick spin. Let it site for 15 mins before using. Recommend 5 µl per 100 µl of any staining buffer for 1 million cells. • For ICC staining Recommend 15 µl per 100 µl of any staining buffer for 1 million cells.