Assay cellulose filter binding

The modified cellulose filter binding assay is based on the tight binding of proteins to this kind of filter material. When a protein-nucleic acid mixture is filtered, proteins are retained on the filter while nucleic acids are washed through. However, nucleic acids are also retained on the filter if they are bound to proteins. Thus, free and protein-bound nucleic acids can be separated [43]. 

The preparation of the cyclic AMP binding protein from bovine muscle has been described [150]. It involves homogenisation, centrifugation, pH 4.8 precipitation and ammonium sulphate precipitation, followed by fractionation on DEAE cellulose. The preparation binds 0.3 pmol of cyclic AMP per ixg of protein and has an enzymatic activity of 24 pmol of P per jxg of protein per 10 min. Over 200 /ig of the protein can be quantitatively adsorbed on a single Millipore filter. The yield of binding protein from 500 to 1000 g of muscle is sufficient for more than 100000 assay tubes. The binding activity is stable for 18 months at -20°C. 

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