I read in most papers that they either use ~4% or ~8% EDTA inPBS solution when mice/rat mandible/long bone is decalcified for immuno purpose. I myself always used 8% for faster process. Last time in the conference, a senior technician said that 8% EDTA is too high. Anyone know what are the differences between 4% and 8%? Will the 4% preserve the ultrastructure or antigens better plz? Thanks a lot!
See my paper on using 4.13% disodium EDTA. The paper with Warshawsky and Moore. It is best for EM because it is neutral buffered and isotonic. HW.
0.33 Molar is both isotonic and safe. But remember to nutralise it using NaOH to pH of 7.2 if you are demineralising for histology purposes; especially if you want to do any immunohistochem on the sections.
I have used 0.5 M EDTA in buffer for many IHC and ISH. Didn't have notable problem.
Can anyone tell why higher conc. EDTA will harm?
See Warshawsky and Moore 1967 for a complete protocol on 4.13% disodium EDTA.
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