Download e-book for kindle: DNA-Protein Interactions: Principles and Protocols by Peter G. Stockley (auth.), Tom Moss (eds.)

By Peter G. Stockley (auth.), Tom Moss (eds.)

ISBN-10: 0896036251

ISBN-13: 9780896036253

ISBN-10: 0896036715

ISBN-13: 9780896036710

Univ. Laval, Quebec, Canada. deals an array of greater than a hundred present and reproducible innovations biochemists and molecular, mobile, and developmental biologists can use to light up DNA-protein interactions. contains protocols for in vivo footprinting. Hardcover, softcover now not on hand.

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Additional info for DNA-Protein Interactions: Principles and Protocols

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Little or no Sp1 binding to its high-affinity binding site (5'-GATCATATCTGCGGGGCGGGGCAGACACAG–3') (48) is usually indicative of a poor quality nuclear extract. Although such an assay is clearly invaluable when crude extracts are obtained from established tissue-culture cells, caution must be observed when extracts are prepared from whole animal tissues because not all organs express Sp1 at the same level (22,47,49). The analysis of crude extracts prepared from whole animal tissues by EMSA is somewhat restricted because of the numerous enzymatic activities, such as proteases and deacetylases, these may contain.

Little or no Sp1 binding to its high-affinity binding site (5'-GATCATATCTGCGGGGCGGGGCAGACACAG–3') (48) is usually indicative of a poor quality nuclear extract. Although such an assay is clearly invaluable when crude extracts are obtained from established tissue-culture cells, caution must be observed when extracts are prepared from whole animal tissues because not all organs express Sp1 at the same level (22,47,49). The analysis of crude extracts prepared from whole animal tissues by EMSA is somewhat restricted because of the numerous enzymatic activities, such as proteases and deacetylases, these may contain.

EMSAs for Analysis of DNA–Protein 27 11. Ethidium bromide is a powerful carcinogen that also possesses a moderate toxicity. Wearing gloves is essential when manipulating solutions that contain this DNA dye. Decontamination of ethidium bromide-containing solutions can be achieved using either hypophosphorous acid or potassium permanganate (see ref. 41 for an overview and detailed protocols). 12. Nearly all vertical electrophoresis apparatus can be used to perform EMSA analyzes. Although gel electrophoresis is performed at room temperature in some EMSA protocols, we recommend 4°C.

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DNA-Protein Interactions: Principles and Protocols by Peter G. Stockley (auth.), Tom Moss (eds.)


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