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Fluorokine E Assay Principle
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Fluorokine E Assay Principle |
Fluorokine E
Step 1
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Step 2
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| Protease-specific antibody (purple) is pre-coated onto a black microplate. The sample (blue) is added and both pro- and active forms of the enzyme are bound to the immobilized antibody. |
A protease-specific peptide substrate (green & purple) is added. The substrate features a fluorophore (FL) and quencher (Q) molecule on opposite sides of the prospective cleavage site. |
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Step 3
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Step 4
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| Active enzyme cleaves the peptide substrate between the fluorophore and the quencher molecules, increasing the distance between them. Energy from the fluorophore is now available as fluorometric signal since the quencher is no longer close enough to absorb it. |
Signal is directly proportional to the amount of active protease bound in the initial step. |
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