BINDING OF BIOTINYLATED DNA TO STREPTAVIDIN-COATED POLYSTYRENE LATEX

被引:32
作者
HUANG, SC
SWERDLOW, H
CALDWELL, KD
机构
[1] UNIV UTAH, DEPT CHEM & FUELS ENGN, SALT LAKE CITY, UT 84112 USA
[2] UNIV UTAH, DEPT HUMAN GENET, SALT LAKE CITY, UT 84112 USA
关键词
D O I
10.1006/abio.1994.1514
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The binding of 5'-end biotinylated DNA fragments was compared between streptavidin (SA)-coated commercial. M-280 magnetic latex particles with a diameter of 2.8 mu m and adsorption-coated polystyrene (PS) latex standard particles whose diameter is 0.272 mu m. Amino acid analysis showed the protein content of the commercial particles to correspond to 4X monolayer coverage, while the adsorption-coated PS particles displayed monolayer coverage, or 8 pmol/cm(2). A fluorescence-based method was developed to quantify the adsorption of FITC-labeled SA, biotin, and biotinylated DNA. The validity of the method was substantiated for the labeled protein by both amino acid analysis and a colorimetric protein assay. While the specific binding of biotin was 0.38 mol per mole of SA on the adsorption-coated 0.272-mu m particles and slightly higher (0.6 mol per mole SA) on the 2.8-mu m particles, the specific binding of the bulky biotinylated 300-bp DNA was more favorable on the smaller particle (0.12 mol per mole SA for 0.272 mu m versus 0.04 mol per mole SA for 2.8 mu m). (C) 1994 Academic Press, Inc.
引用
收藏
页码:441 / 449
页数:9
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