Analysis on musculature proteome of rat fetus by two-dimensional gel electrophoresis

  Objective   To establish protocol of two-dimensional Gel Electrophor esis for analysing the proteome of mus-culature of Wistar rat fetus.

  Methods   Proteins extracted from tibia-fibulae musculature isolated from 21-day ratfetus were loaded on immobile pH gradient gels to process the first dimension isoelectric focusing electro phoresis and then transfer to SDS poly acrlamide gel for the second dimension electrophoresis.Following the electrophoresis Coomassie Brilliant Blue staining or silver staining was performed, and the images were scanned and analyzed with Image Master 5.0 software package.One set or protein spots matching from three separate gels were cut for MALDI-TOF mass spectrometry analysis.

  Results   Images with good reproducibility of the proteome of musculature were obtained after tow dimensional electrophoresis.The reproducibility of 2D gelelectr ophoresis was verified through comparing of peptide mass fingerprints form matching protein spots of three separate gels which sug gested a same protein(light chain 1 of skeletal muscle myotonin).

  Conclusion   The utilization of 2D electrophoresis can satisfactorily separ ate muscular proteins from ratembryos and achieve a high resolution and good repr oducibility for protein identification.