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.
Inomata
Y
Y
Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama, Japan.
Wada
T
T
Handa
H
H
Fujimoto
K
K
Kawaguchi
H
H
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