I noticed that almost all of the IP buffer containing magnesium ions, but I don't know the exact function of Mg2+ in the cell lysis process. Could anyone provide the answer of it? To my knowledge, Mg2+ contributes to chromosome condensation. What consequences would it be if there is no Mg2+?
It contributes to the elimination of DNA from cell lysates after addition of of protein G or A conjugated resins (substances) when you separate the supernatant fraction from the bound antibody-protein complex by centrifugation or when you collect the complex using magnetic beads.
I agree with Vehary. Additionally, note that when membranes are broken by TRIS, compartmentalization in the solution is compromised. Mg2+ then bind to DNA and protect it against DNase proteins that are now in direct contact with the DNA. DNAse, as a protein in itself, is spared a degradation activity and can also be precipitated, if it is the target protein. Mg2+ also mimic the ionic interactions that might have occurred during and/or post lysis.
Hi Shirley, my thinking is that Mg++ complexes with and condenses the phosphate backbone of DNA so it does not form the DNA sticky "snot" when you lyse cells with detergents alone.
Try lysing cells with just detergents and you will see how snotty the lysate will be.
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