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Primary osteoblasts were fixed in 70% ice-cold ethanol for 1 h and rinsed with distilled water. For Alizarin red staining, fixed cells were stained with 40 mM Alizarin Red S (pH 4.2) for 10 min with gentle agitation. The level of Alizarin Red S staining was observed under light microscopy. An ALP staining kit (86R-1KT, Sigma-Aldrich) was used according to the manufacturer's instructions.
Copyright and License information: ©2016 Auh et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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