Detection and analysis of neutral endopeptidase from tissues with substrate gel electrophoresis |
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Authors: | J Sullivan A R Johnson |
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Institution: | Department of Biochemistry, University of Texas Health Center, Tyler 75710. |
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Abstract: | Neutral endopeptidase from human or bovine tissues retains enzymatic activity following electrophoresis and immobilization in polyacrylamide gels. Infiltration of the gel with a fluorogenic substrate permits identification of the active enzyme by fluorescence associated with a distinct protein band. This technique both separates and identifies the enzymatically active species from a crude cell membrane fraction or from partially purified extracts that contain contaminating proteins. Enzymatic activity is quantitated by photographing the fluorescent bands and scanning the negatives with a laser densitometer. Because as little as 25 ng of enzyme can be detected by this method, it could be used where the amount of material is limited. |
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