Cloning of an antibody binding DNA sequence: pitfalls of DNA/protein immunoprecipitation reactions.
This paper presents evidence of cross-reactivity of a normal genomic DNA sequence to a number of antibodies raised against common proteins. This DNA fragment was inadvertently isolated during DNA-protein immunoprecipitation cloning experiments and was found to bind not only to the antibody used in the immunoprecipitation experiment, but to several different antibodies in gel-shift assays. This data demonstrates the ability of certain DNA sequences to bind non-specifically to antibodies and highlights the danger of DNA/antibody binding which can produce artifacts. Possible means of overcoming these reactions are discussed.
Department of Medical Biochemistry, University of the Witwatersrand Medical School, Johannesburg, South Africa.
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Weitzmann
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Savage
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D001665:Binding Sites; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D011233:Precipitin Tests / Q000379:methods*; D011485:Protein Binding
D000906:Antibodies; D004247:DNA