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A microscale analytical batch chromatographic method for detecting soluble viral DNA-binding proteins in crude
1Department of Biological Sciences, Illinois State University, Normal 61761.
Abstract:
We describe a micro-method for determining the presence in crude cellular extracts of soluble proteins which can bind to immobilized DNA, using the DNA-binding protein of human adenovirus as an example. Batch chromatography of radiolabeled proteins is performed in microcentrifuge tubes containing 50 microliters packed volume of commercially available denatured calf thymus DNA-cellulose. Eluted single-stranded DNA-binding proteins are then visualized by fluorography following gel electrophoresis. The batch procedure gives yields of adenovirus DNA-binding protein which are comparable to those obtained with a mini-column of similar adsorbent volume. The scale of the procedure makes it convenient for simultaneously analyzing multiple samples.