Use of GFP as a reporter for the analysis of sequence-specific proteases.


This unit describes a rapid fluorescent assay for sequence-specific proteases. A recombinant His-tagged substrate-GFP fusion protein containing the sequence-specific protease-recognition sequence is used as substrate. Batch metal-chelate chromatography separates uncleaved substrate-GFP fusion protein from GFP released by proteolysis and proteolytic activity is determined by measuring the fluorescence of GFP remaining in solution.

Publication Title

Current protocols in protein science / editorial board, John E. Coligan ... [et al.]

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