[Establishment of two-dimensional electrophoresis system of caudal gland].

Objective: To establish an effective separation system of 2-DE for the proteome of caudal gland,and provide foundation for revealing the mechanisms of histological development and pharmacological activities.Method: The total proteins of caudal gland were extracted by TCA/acetone precipitation,phenol extraction/methanol-ammonium acetate precipitation and trizol-base method respectively and separated by immobilized pH gradient(IPG) strips prior to SDS-PAGE.Loading protein sample size and isoelectric focusing conditions were optimized.The gels were stained with Coomassie brilliant blue,scanned and then analyzed using PDQuest 8.0 analysis software.Result: The total proteins of caudal gland extracted by trizol-base method were the highest quality and could meet the needs of 2-DE.With 300 μg of proteins loaded on 7 cm pH 3-10 IPG strip followed by isoelectric focusing programⅡ,a satisfying 2-DE profiles were obtained.The total number of disticted protein spots was 209 with the optimized system.Conclusion: A well-resolved 2-DE patterns of caudal gland were obtained by this optimized system.This method could be applied to prepare other similar tissue sample and 2-DE studies.