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The TRAP Stain is used for the staining of tartrate-resistant acid phosphatase in osteoclasts. Bone mass is controlled by the balance between the activity of osteoblasts and the activity of osteoclasts. Alkaline phosphatase and tartrate-resistant acid phosphatase are used as markers for osteoblasts and osteoclasts, respectively.
Formalin fixed, paraffin tissue sections.
Mineralized bone MUST be decalcified using EDTA.
Acid-decalcifiers inhibit enzymatic staining.
1) Place TRAP Staining Solution Mix in staining dish and pre-warm to 37°C in waterbath.
2) Deparaffinize slides and rehydrate through graded ethanols to distilled water.
3) Place slides in pre-warmed TRAP Staining Solution Mix and incubate at 37°C for 30 mins. or until control is developed.
4) Rinse in distilled water.
5) Counterstain with hematoxylin for 30 sec. and rinse quickly in distilled water.
6) Dehydrate quickly through graded alcohols, 5 seconds each, clear in Xylene and mount.
Osteoclasts: red violet