Preparation of DNA-carrying affinity latex and purification of transcription factors with the latex.
We have developed DNA-carrying latex particles for the separation and purification of transcription factors. These particles consist of styrene (St), glycidyl methacrylate (GMA) and divinylbenzene (DVB). It was confirmed that the ethanolamine-treated surface of these particles suffered no nonspecific adsorption of proteins. To the latex particles sequence-specific DNA oligomers were immobilized via covalent coupling. A transcription factor, E4TF3, was efficiently purified to homogeneity using the latext particles. In contrast, the purification using DNA-carrying Sepharose gel yielded poor results. Compared to DNA-carrying Sepharose gel, the latex particles exhibited several times higher efficiency in the purification of E4TF3 from the crude nuclear extract.
Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama, Japan.
Y
Y
Inomata
T
T
Wada
H
H
Handa
K
K
Fujimoto
H
H
Kawaguchi
D000327:Adsorption; D001483:Base Sequence; D004852:Epoxy Compounds / Q000032:analysis; D007840:Latex; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D012685:Sepharose; D014157:Transcription Factors / Q000302:isolation & purification*
D004852:Epoxy Compounds; D007840:Latex; D009841:Oligonucleotides; D011506:Proteins; D014157:Transcription Factors; D012685:Sepharose