Azure B (Azure B chloride) is a cationic dye and the major metabolite of Methylene blue. Azure B is a selective, and reversible inhibitor of MAO-A (IC50s: 11 and 968 nM for recombinant human MAO-A and MAO-B).

MAO-B:968 nM, MAO-A:11 nM

Azure B (4-30 mg/kg; i.p.; once) reduces immobility in the forced swim test [1].

Instructions
1. Solution configuration
1. Configuration of mother liquor: Use sterile water or PBS to configure MHI-148 into 1-10mM mother liquor and store it at -20℃ or -80℃. Please aliquot it to avoid repeated freeze-thawing.
2. Configuration of working fluid: When in use, dilute the MHI-148 mother liquor into a working fluid that meets the requirements and adjust according to the experimental requirements.
2. Blood smear staining:
1. Prepare an Azure B staining solution, usually by dissolving Azure B chloride in buffer or solvent.
2. Apply the staining solution to the prepared blood smear.
3. Allow the smear to incubate in solution for a few minutes to promote cell staining.
4. Rinse the smear thoroughly with water to remove excess dye and then air dry.
5. Check the stained smear under a microscope and analyze the morphology of blood cells.
3. Monoamine oxidase (MAO) inhibition:
1. Introduce Azure B into in vitro experiments, the inhibitory effect is usually determined using recombinant human MAO-A and MAO-B.
2. The IC50 value at a specific experimental setting can be determined through dose response studies.
3. In addition, the antidepressant effect of Azure B can be evaluated in animal models through behavioral experiments (such as forced swimming experiments, tail suspension experiments).

The above information is based on published literature. Experimental procedures should be appropriately modified to meet specific research demands.

DMSO: Insoluble