[Histonet] EDTA decalcification solution

[Rene J Buesa]

Fawn:
  This is how I used to prepare it:
   
  EDTA --- 250g
  Dist. water ---1,750 mL
   
  Add the EDTA to the water in a hot plate with constant agitation (teflon covered magnet). When all the salt is dissolved the solution will have a "milky" appearance. Leave the solution reach room temperature and determine pH (ideally with a pH-meter). The solution will be acid (<pH7.0) but it should be neutral.
   
  To get it to neutral (pH7.0) add sodium hydroxide in pellets, small amount every time, with agitation. Usually you will need abbout 25g of NaOH to reach pH7.0, but that amount can vary somewhat. 
  Once the solution is at pH7.0 it will no longer be milky; it will be clear (this is also an indication that it is at pH7.0)
  René J.

Fawn Jones <fawn <@t> cs.cmu.edu> wrote:
  Hi everybody,

I am getting ready to decalcify some mouse femurs for immuno staining. 
I know I have to use an EDTA decalcifier, but I do not know how to make 
it. Does anybody have a protocol on this that they would be willing to 
share?

Thanks in advance
Fawn

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