Apotracker Red

Operating Instructions
(The following is a recommended experimental protocol for guidance only; please adjust it according to your specific needs).

1. Stock Solution Preparation
1.1 Solvent: Most dyes are usually dissolved in organic solvents, such as anhydrous DMSO.
1.2 Concentration Recommendation: High-concentration stock solution of 1-10 mM.
2. Working Solution Preparation
2.1 Diluent: Serum-free medium or PBS is commonly used.
2.2 Working Concentration: 150 nM.
2.3 Notes: Adjust the working concentration of the dye according to actual conditions, and prepare it immediately before use.

3. Staining Procedures
3.1 Sample Types: Suspension cells (Jurkat cells, human neutrophils, cancer cell lines)^[1]
3.1.2 Suspension cells: No centrifugation is required for wash-free staining.
3.2 Incubation Conditions: Incubate with 150 nM Apotracker Red for 10-15 min.
3.3 Washing Step: No washing step is needed; adopt the wash-free staining method.

4. Detection and Analysis
4.1 Instruments: Fluorescence microscope, flow cytometer, multiphoton microscope.
4.1.1 Single-photon excitation/emission: 530 nm/~600 nm, 561 nm/610 nm; two-photon excitation/emission: 1070 nm/620 nm.
4.2 Result Analysis:
4.2.1 Changes in fluorescence intensity: Apoptotic cells exhibit bright red fluorescence; viable cells show only extremely weak or no fluorescence.
4.2.2 Fluorescence localization: Red fluorescence localizes on the surface of apoptotic cells.
4.2.3 Changes in fluorescence state: The fluorescence is in the OFF (no fluorescence) state in viable cells, and switches to the ON (red fluorescence) state in apoptotic cells.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.