Acid phosphatase assay cell type - human periodontal ligament cells

Acid phosphatase detection heavily relies on determining the concentration of tartrate-resistant acid phosphatase (TRAP) in the sample. Hence, sample preparation is very crucial and it should be done strictly as per kit manufacturer instructions to avoid any inconsistency and poor sensitivity

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Protocol tips
PDL cells were fixed by immersion in a citrate-acetone-formaldehyde fixative solution for 5 min. The alkaline-dye mixture was prepared by mixing 1 mL Sodium Nitrite Solution and 1 mL of a fast red violet alkaline solution dissolved in 45 mL of distilled water and 1 mL of Naphtol AS-Bl alkaline solution. Surfaces were then placed in an alkaline dye mixture solution for 15 min protected from light followed by rinsing in deionized water.
Downstream tips
All images were captured on a Wild Heerbrugg M400 ZOOM Makroskop (WILD HEERBRUGG, Heerbrugg, Switzerland) at the same magnification and light intensity and imported into Image J software (NIH, Bethesda, MD, USA). Image segmentation (Thresholding) was used to generate percent stained values for each field of view.
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