| 摘要 |
A simple and quick protocol for chemical treatment, enzymatic or chemical digestion, and subsequent identification of proteins on protein chip arrays is disclosed, together with kits therefor. The chemical treatment comprises denaturation, reduction and alkylation while enzymatic digestion encompasses deglycosylation, dephosphorylation, and digestion by various proteases. Digestion is also accomplished by various chemicals that are known to induce proteolysis. All reactions are carried out sequentially on chip. Subsequent peptide mass fingerprinting or product ion searches allow the identification of specific peptides which can be correlated to proteins. The method of the present invention can be applied to the analysis of biological samples such as urine and plasma to identify biomarkers in diseased states. The methods of the present invention allow complete on chip treatment, which can be used for rapid protein identification and structural characterization of heavily posttranslationally modified proteins.
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