TRAP for bone osteoclasts

From:Gayle Callis <>

There is an excellent publication on TRAP staining  is J Histochem Cytochem
by Andersson G and S C Marks Jr.  Titled tartrate resistant acid ACTapse as
a cytochemical marker for osteoclasts.  

The following publication Eggert and Germain, Stabel acid phosphatase: I.
Demonstration and distribution, Histochemistry 66:307-317, 1980 discusses
the use of rapid acidic buffer decalcification for demonstration of acid
phosphatase.  Buffered  formic acid (with sodium formate or sodium citrate)
is an acidic buffer.

An excellent article on technic in Journal of Histotechnology.  Parkinson I
et al, The histochemical localization of osteoclasts in EDTA decalcified

EDTA may take longer but you must protect enzyme activity from damage by
acid pH, and you can control the pH level better with EDTA. Parkinson's
method is certainly worth a try for speed.  

Many people use TRAP staining on GMA and PMMA plastics embedded bone, also
undecalcified bone frozen sections, not the easiest to do.  

The TRAP protocol used by SC Marks for deparaffinized sections was Barka
and Andersons, and also used a modified method for tartrate resistant
AcATPase from Wachstein and Meisel, Am J Clin Pathol 27:13, 1957.  Marks
has many publications on TRAP staining, and his results are excellent, with
wonderful photos and the above reference gives recipe for substrate.  

Good luck on staining success


Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
Bozeman MT 59717-3610

406 994-6367
404 994-4303 (FAX)

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