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Nitrocellulose Filter Binding Assay

Information on nitrocellulose filter binding assay.

Nitrocellulose Filter Binding Assay

Please refer to the diagram below

a) Double Stranded DNA

The double stranded DNA is end labeled (pink) and passed through a nitrocellulose filter (yellow). Then radioactivity on the filter paper and filtrate is monitored by liquid scintillation counting. None of the radioactivity sticks to the filtedr, indicating that double-stranded DNA does not bind nitrocellulose. Single stranded DNA, on the other hand binds tightly.

b) Protein

First we label a protein (orange) and filter it through nitrocellulose. The protein binds to the nitrocellulose.

c) Double-Stranded DNA- Protein Complex

We mix an end labeled double stranded DNA (pink) with an unlabeled protein (orange) to which it binds to form a DNA-protein complex. Then we filter the complex through nitrocellulose. The labeled DNA now binds to the filter because of the association with the protein. Thus, double stranded DNA protein complexes bind to nitrocellulose, and this provides a convenient assay for association between DNA and protein.

nitrocellulose filter binding assay

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