I am searching for a robust method to be able to quantify at the same time amounts of protein, nucleic acid and the ribonucleoprotein complexes in a defined nucleic acid/protein binding assay. What would be your method of choice? Regards and many thanks in advance for your consideration.
Benyamin
Dear Benyamin,
It is very interesting question and its answer is not easy. Some methods are general for a kind of molecule and some specific for a special molecule. e.g binding of Cu++ to proteins is a general but binding BCG to Albumin is specific for albumin.
Anyway I think a robust method is photometric methods, but it has its limitation too.
In other words it is very difficult finding a perfect and robust method for all three kinds mentioned molecules.
In photometric method, e.g absorbance at 260nm and 280nm and their ration may help us. It cannot discriminate among protein/ nucleic acid mixtures and nucleoproteins (also 230nm for organic compounds).
This method will limit by absorbance of proteins, peptides and amino acids and also the similar limitation for nucleotides, oligonucleotides and poly nucleotides.
Anyway, for determination of all components in a sample at the same time, I suggest you using photometric assay and matrix analyzing formula (chemometry or biochemometery methods).
This is robust and no need to separating components and determination separately.
the attached may help you too.
best
mehdi
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