Record number :
Title of article :
DNA–support coupling for transcription factor purification: Comparison of aldehyde, cyanogen bromide and N-hydroxysuccinimide chemistries
Author/Authors :
Chockalingam، نويسنده , , Priya Sethu and Gadgil، نويسنده , , Himanshu and Jarrett، نويسنده , , Harry W، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2002
Pages :
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Abstract :
Purification of transcription factor IIIA on internal control region DNA coupled to aldehyde-silica is described and compared with purification on cyanogen bromide-activated Sepharose and Bio-Rad Affi-Gel-10. The Affi-Gel support results in mixed-mode chromatography; both ion-exchange and affinity modes contribute. Coupling DNA to aldehyde-silica is advantageous in that it has no ion-exchange properties and performs as well as DNA coupled to CNBr-activated Sepharose. Purification of lac repressor on aldehyde-silica, and CAAT enhancer binding protein on Affi-Gel also shows the advantages of a neutral support and the disadvantages of mixed-mode chromatography for transcription factor purification. Aldehyde-silica couples to alkylamines and to the amines of adenine, guanine, and cytosine nucleoside bases. Reaction occurs with either single- or double-stranded DNA, although it is less efficient with the latter. Overall, the results demonstrate that predominantly neutral coupling chemistries, such as aldehyde or CNBr-mediated coupling, have distinct advantages for transcription factor purification. Since the CNBr chemistry has not yet been applied to silica supports, aldehyde-silica coupling is currently the most attractive method for DNA affinity HPLC.
Keywords :
DNA , Proteins
Journal title :
Journal of Chromatography A
Journal title :
Journal of Chromatography A
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Link To Document :