Does the degree of denaturation affects the digestion processalong with the lc-ms/ms result?
If so then how does it affects?
Hi,
In general, the more a protein is denatured, the better it can be digested, as more potential proteolytic cleavage sites become available once a protein unfolds.
I'm not sure if I understand your question, but a protocol referred to as 'pellet digestion' has been developed to quickly and efficiently digest proteins/protein mixtures.
Please refer to J Proteome Res 2009 8(6):2838-50 for a proteomics application of pellet digestion, or to Bioanalysis 2012 4(24):2887-96 for an application describing bioanalysis of a monoclonal antibody.
Hope this helps!
Hi Anjali,
Protein gets denatured by organic solvent precipitation won't affect the digestion efficiency. You can re-dissolve it in ammonium bicarbonate buffer to perform the in-solution digestion with no problem.
Hope it helps :)
Thank you for the suggestion Chi-Chi Chou.
I usually re-dissolve it in ammonium bicarbonate.
But I'm facing some issues with the acetone precipitation method.
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