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Alizarin Red S staining and determination of cetylpyridinium chloride (CPC)
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MC3T3-E1 cells were fixed with 4% paraformaldehyde for 30 min and then incubated with Alizarin Red S (sigma Aldrich) for 10 min. After washing with deionized water, calcium deposition was observed by optical microscope. 10% cetylpyridinium chloride (CPC, sigma Aldrich) was used for quantification, and the absorbance value was detected at 562 nm wavelength with a microplate meter (BMG Labtech, Germany).
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