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From:Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of Marvin Hanna)
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EDTA, disodium 10% in PBS, some use water, but I like to maintain the cells

EDTA alone (no sodiums attached) 10%, same as above

EDTA tetrasodium salt, 14%, dissolve in 800 mls water, adjust the pH with
glacial acetic acid, not HCl! You must adjust the pH of this tetrasodium
salt, as it runs above pH 8, around 9, and that is not good for alkaline
sensitive protein linkages.

pH range is 7 to 7.4, like the 7.4

Use a good volume to tissue ratio, 20 to 1, I always liked more (is

Be patient, suspend the specimen in center of this solution, to let EDTA
touch all surfaces of bone.  It can be left over a weekend or longer, but
do change it a couple of times a week, depending on size or how many bones
are in the solution.

Test for completetion of decalcification, rinse EDTA out of bone or a ppt
will form in bone when it contact the first alcohol

Good luck

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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