Protein purification from polyacrylamide gels by sonication extraction

被引:78
作者
Retamal, CA
Thiebaut, P
Alves, EW
机构
[1] Univ Estadual Norte Fluminense, CBB LQFPP, Ctr Biociencias & Biotecnol, Rio De Janeiro, Brazil
[2] Univ Fed Rio de Janeiro, DBM ICB, Dept Bioquim Med, Rio De Janeiro, Brazil
[3] Univ Chile, Fac Med, Santiago 7, Chile
关键词
gel electrophoresis; proteins; protein purification; native proteins; sperm proteins;
D O I
10.1006/abio.1998.2977
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A proteinpurification procedure using sonication extraction from polyacrylamide gels (PAGE), which involves identification of a particular protein band and its excision, homogenization, sonication, and subsequent passage through a Sephadex G-25 minicolumn, is reported. Our results show a high degree of recovery regardless of the nature of the protein (soluble or membrane bound) or the characteristics of the gel (SDS-PAGE, native gels, or Tricine-SDS-PAGE). The percentage of recovery was dependent on the protein concentration applied in the gel. This technique is fast, gives high yield, provides good resolution, and can be used without any specialized equipment. The method was tested with a wide variety of membranes and soluble proteins and was found to give good results and to be applicable to different studies. The amino acid sequence of one of the purified proteins (Rf 0.45 stallion ejaculated sperm protein) was determined successfully. (C) 1999 Academic Press.
引用
收藏
页码:15 / 20
页数:6
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