To measure osteoclast markers, the section was staining by TRAP staining (Sigma-aldrich, catalogue no. 386A, Saint Louis, MO, USA) with according to the instructions of the manufacturer. Briefly, we deparaffinized and dehydrated the samples by xylene and alcohol. Samples were covered with fixative buffer for 30 s and then rinsed in deionized water. Naphthol AS-BI phosphoric acid solution and Fast Garnet GBC base solution (Sigma-aldrich, catalogue no. 386A, Saint Louis, MO, USA) were used for TRAP staining with cover samples for 2 h at 37 °C. The samples were then washed with pure water and counterstained with hematoxylin. Each section was observed and recorded by upright microscope with image capture system. The calculating average of TRAP-positive cells was analyzed under 200× magnification in six different fields.
Naphthol as bi phosphoric acid solution
Manufactured by Merck Group
Sourced in United States
Naphthol AS-BI phosphoric acid solution is a laboratory reagent used for the detection and visualization of various enzymatic activities in biological samples. It is a colorless to pale yellow liquid that serves as a substrate for the detection of enzymatic activity, particularly phosphatases, through the release of a colored product.
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Lab products found in correlation
2 protocols using naphthol as bi phosphoric acid solution
1
Quantifying Osteoclast Activity via TRAP Staining
2
Quantifying Osteoclasts During Orthodontic Tooth Movement
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To estimate the number of osteoclasts during OTM, tartrate-resistant acid phosphatase (TRAP) staining was performed. Sections were incubated in acetate buffer containing naphthol AS-BI phosphoric acid solution, Fast Garnet GBC base solution, sodium solution, and tartrate solution (Sigma-Aldrich, St. Louis, MO), and then counterstained with hematoxylin. The number of TRAP-positive multinucleated cells containing more than three nuclei was determined by microscopy. Three sections of the mesial side of the palatal root at 40, 80, and 120 µm from the bifurcation in each mouse were evaluated and four biological replicates were analyzed, and the number of osteoclasts was expressed as the average of these three sections. UPM Axio Phot2 (Carl-Zeiss, Jena, Germany) was used for TRAP staining.
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