Protein purification from polyacrylamide gels by sonication extraction

A proteinpurification procedure using sonication extraction from polyacryla mide gels (PAGE), which involves identification of a particular protein ban d and its excision, homogenization, sonication, and subsequent passage thro ugh 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 b ound) or the characteristics of the gel (SDS-PAGE, native gels, or Tricine- SDS-PAGE). The percentage of recovery was dependent on the protein concentr ation applied in the gel. This technique is fast, gives high yield, provide s good resolution, and can be used without any specialized equipment. The m ethod 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. T he amino acid sequence of one of the purified proteins (Rf 0.45 stallion ej aculated sperm protein) was determined successfully. (C) 1999 Academic Pres s.