Research on Two-Dimensional Electrophoresis Conditions for Proteomic Analysis of Pteridium aquilinum

Abstract:

In order to establish two-dimensional electrophoresis technology system of total proteins of Pteridium aquilinum,2-DE maps was compared with protein extraction method,loading amount and pH ranges of IPG strips The results showed that phenol extraction protocol combined with TCA/ acetone was more favorable for total protein extraction from fresh Pteridium aquilinum.A high quality 2-DE map with more protein spots,better dispersion,clearer background and higher protein point resolution were acquired by using the following optimized parameters:loading 1200μg protein sample on 17cm IPG strip with pH 5-8,and finally detecting proteins with Coomassie brilliant blue G250 staining.Under above experimental conditions,the total number of protein spots is 719,this experimental results provides a basis for further study of post-harvested Pteridium aquilinum proteomics.