What is the role of EDTA-Na2 in sodium phosphate buffer? using for NADPH oxidoreductase assay?
Hello Shurog,
EDTA sodium salt acts as a chelating agents and sequesters all the divalent metal ions like Ca2+ and Mg 2+ , which can otherwise oxidize certain groups like the -SH to -SHO.
It is also very essential to maintain an optimum concentration of EDTA in your buffer system as it may have a negative influence on the enzymatic activity.
EDTA is a synthetic amino acid which chelates divalent cations and is therefore used in various buffers. Most enzymes that synthesize or modify nucleic acids (e.g. polymerases, ligases, kinases, nucleases) are Mg2+ dependent. The addition of EDTA is a expedient way to stop these reactions, and helps in the removal of the metal cofactors (typically Mg++ required for activity of DNAses and other DNA damaging enzymes).
Many nucleaes which degrade DNA and RNA require divalent cations (Mg++, Ca++, or Mn++) to enable their activity.
EDTA chelates (binds) divalent catiors, thereby rendering many nucleases inactive.
Therefore, when purifying (or otherwise manipulating DNA or RNA) in procedures that don’t require divalent cations, including EDTA is recommended to avoid enzymatic degradation of DNA and RNA by contaminating nucleases.
-paul r walsh
Divalent cations such as Mg2+, Zn2+, and Cu2+ is bind with sodium EDTA . with this you no free divelent ions present in your protocol, its work as a metalloproteases to.
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