Summary
The strong interaction of chelated nickel ions with a hexahistidine peptide was utilized to identify hexahistidine fusion proteins in crude cellular lysates. The technique involves a Ni2+-nitrilotriacetic acid derivative labelled with alkaline phosphatase. The new reagent was used to probe for hexahistidine fusion proteins after SDS-PAGE electrophoresis and blotting onto nitrocellulose. The minimal amount of fusion protein which could be detected with this method was 0.1 μg.
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Piesecki, S., Hochuli, E. Method to identify hexahistidine fusion proteins in crude cellular lysate. Biotechnol Tech 8, 39–44 (1994). https://doi.org/10.1007/BF00207631
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DOI: https://doi.org/10.1007/BF00207631