AM4-66

Operating Instructions
(The following is a recommended experimental protocol for guidance only, and adjustments are required based on your specific needs).

1. Stock Solution Preparation
1.1 Solvent: For most dyes, organic solvents such as anhydrous DMSO are commonly used for dissolution.
1.2 Concentration Recommendation: It is generally recommended to prepare a high-concentration stock solution with a concentration of 1-10 mM.

2. Working Solution Preparation
2.1 Diluent: Serum-free medium or PBS is usually used. Proteins and esterases in serum may interfere with staining or cause dye hydrolysis.
2.2 Precautions: Adjust the working solution concentration as needed; prepare and use it immediately.

3. Staining Procedure
3.3 Sample Type: Adherent cultured ventricular cardiomyocytes from adult rats, rabbits and humans.
3.2 Incubation Conditions: Use AM4-66 as a cell membrane surface dye, applied sequentially after fluorescent dextran labeling.

4. Detection and Analysis
4.1 Instrument: Confocal microscope.
4.2 Result Analysis: AM4-66 labels the cell membrane surface; Dextran-positive structures inside cells not stained by AM4-66 indicate that transverse tubule membranes have detached from the cell surface. For sequential labeling, the overlap rate of dextran-positive voxels and AM4-66-positive voxels is approximately 11.8%, while the overlap rate for simultaneous labeling is approximately 47.6%.

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