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Biochemistry
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Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
 

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Article DOI: 10.3791/58824-v 19:23 min January 16th, 2019
January 16th, 2019

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Here, we present the detailed procedure of a recently developed protease assay platform utilizing N-terminal hexahistidine/maltose-binding protein and fluorescent protein-fused recombinant substrates attached to the surface of nickel-nitrilotriacetic acid magnetic agarose beads. A subsequent in-gel analysis of the assay samples separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is also presented.

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Keywords: Recombinant Fusion Proteins Fluorescent Protease Assay In-gel Renaturation HIV-1 Protease High-throughput Screening Fluorescence-based Assay Protein Expression Cleavage Site BL21(DE3) Cells Plasmid Construction
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