I am trying to extract the protein content of extracellular bone matrix.
After milling the frozen bone in nitrogen liquid, I have used RIPA buffer and HALT to extract the protein content, but the kit also provides EDTA and there is not any indication if its use is recommended or not, only it isn't suggested to use in some protein purification methods...
So, what's is the role of EDTA in protein purification? Is there any difference between use or not use EDTA?
Very often EDTA is employed as a cation chelating agent that removes essential cations required for proteases. That way, EDTA inactivates proteases and the purification/extraction process proceeds with low or negligible proteolytic degradation.
Very often EDTA is employed as a cation chelating agent that removes essential cations required for proteases. That way, EDTA inactivates proteases and the purification/extraction process proceeds with low or negligible proteolytic degradation.
When you break the cells, you release metalloproteinases. EDTA captures the divalent metal ion in their active center, thus deactivating them.
Adrian Velazquez-Campoy Achim Recktenwald can you tell me what is the function of with or without EDTA extracellular protein extraction ?
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