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Mineralization assay by alizarin red S (ARS) staining

AM Arnab Mahato
MD Munmun De
PB Promita Bhattacharjee
VK Vinod Kumar
PM Prasenjit Mukherjee
GS Gajendra Singh
BK Biswanath Kundu
VB Vamsi K. Balla
SN Samit Kumar Nandi
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The mineralization study was assessed by using the ARS staining dye. In brief, prior to washing thrice with PBS, MG-63 cells loaded constructs were fixed in ice cold 70% ethanol for 1 h. Next, constructs were again washed with distilled water and stained with ARS (40 mM) for 20 min at RT. At the end, the stained constructs were viewed under optical microscope after rigorous washing with distilled water. The principle of the study is to observe the mineralization which occurs due to binding of ARS with calcium salts. This can be qualified by imaging (mentioned above) and quantified by reading the absorbance at 540 nm in microplate reader prior to desorbing the stain by using 10% cetylpyridinium chloride for 1 h.

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