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Rationale
Testing of the Autodigestion Hypothesis requires identification of the digestive enzymes involved and quantitative measurements of their activity.
We developed tests for pancreatic chymotrypsin, trypsin, elastase, thrombin , and others.
The Protocol in a Nutshell
(A) A finger prick of whole blood mixed with a charge-changing, protease-specific peptide substrate. After proteolytic cleavage, the substrate splits into a positively charged fraction with a fluorescent beacon;
(B) Placement on a well above the gel;
(C) Electrophoretic displacement of fluorescent, positively charged fragments into the gel and its digital intensity measurement.
Past Applications
A single high-calorie meal results in elevations of pancreatic protease activity in the systemic circulation of healthy controls as well as diabetics.
Digestive enzyme activity and protein degradation in plasma of heart failure patients.
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