[Histonet] Re: TRAP staining on formic acid decalcified bone reference

[koellingr <@t> comcast.net]

Hi all, as usual Gayle was right on.  Use a buffered (more gentle) formic acid; not just formic acid per se of any water diluted concentration.  For end point testing, critical, we used a radiograph machine instead of chemical endpoints which is also fine;we just had access to a lot of equipment. 
Ray 
Washington 

----- Original Message -----

From: "Gayle Callis" <gayle.callis <@t> bresnan.net> 
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, April 2, 2015 3:12:55 PM 
Subject: [Histonet] Re: TRAP staining on formic acid decalcified bone        reference 

The reference within a reference from Ray is A Chimeric Form of 
Osteoprotegerin Inhibits Hypercalcemia and Bone Resorption Induced by IL-1β, 
TNF-α, PTH, PTHrP, and 1,25(OH)2D3 .   Sean Morony et al .  J Bone Mineral 
Res V 14, pp 1478-1485.   

  

However, the formic acid decalcification method is not described in detail 
and merely says "formic acid" but whether this is buffered formic acid or 
just dilute formic acid in water only is not stated.     Ray might elaborate 
on what specific formic acid recipe he used as many in research don't always 
use buffered formic acid decalcifiying solutions.   

  

I would assume Morony et all used a buffered formic acid with either sodium 
formate or sodium citrate and controlled so as to not overexpose TRAP to 
acids longer than necessary.   One publication,  i.e.,  Eggert and Germain. 
Stable Acid Phosphatase I. Demonstration and Distribution.  Histochem 66, pp 
301-317, 1980) discussed in detail the  rapid demineralization in acidic 
buffers i.e. buffered formic acid for staining of stable forms of acid 
phosphatase.   

  

I have both of these publications on file and will forward privately.   

  

I would err on the side of using a buffered formic acid with either sodium 
formate or sodium citrate for doing this and use decalcification endpoint 
testing to avoid over exposure to acid i.e. over decalcification.     

  

Take care 

  

Gayle M. Callis 

HTL/HT/MT(ASCP) 

  

_______________________________________________ 
Histonet mailing list 
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet