Protein Purification from Polyacrylamide Gels by Sonication Extraction

A proteinpurification procedure using sonication extraction from polyacrylamide gels (PAGE), which involves identification of a particular protein band and its excision, homogenization, sonication, and sub- sequent passage through a Sephadex G-25 minicol- umn, is reported. Our results show a high degree of recovery regardless of the nature of the protein (solu- ble 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 pro- tein 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 re- sults 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 deter- mined successfully. © 1999 Academic Press