Alizarin Complexone (Alizarin-3-methyliminodiacetic acid) is an inhibitor of Rous-associated virus 2 reverse transcriptase. Alizarin Complexone can be used as a calcium-tracer and a chelating agent for biological studies.

I. Solution preparation
1. Mother solution preparation: Dissolve Alizarin Complexone in DMSO to obtain the required mother solution, usually 1-10mM.
2. Working solution preparation: Dilute the mother solution with PBS/DMEM/H2O to the required working solution, usually with a concentration range of 1-10μM.
II. Calcium ion detection
1. Add Alizarin Complexone working solution to the cell culture medium and incubate for a certain period of time to allow it to enter the cells and bind to calcium ions in the cells.
2. Use a fluorescence microscope or fluorescence spectrometer to detect the fluorescence signal in the range of 550-600 nm.
3. According to the intensity of the fluorescence signal, calculate the change in calcium ion concentration.
II. Reverse transcriptase inhibition experiment:
1. Dissolve Alizarin Complexone in a suitable solvent, usually at a concentration of 1-10 μM.
2. Mix the dissolved Alizarin Complexone with the RCV-2 reverse transcriptase solution.
3. Evaluate the inhibitory effect of Alizarin Complexone on reverse transcriptase activity by measuring reverse transcriptase activity (e.g., by radiolabeling or changes in fluorescence signal).