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DIGE荧光差异凝胶电泳结合MALDI质谱分析

来源:奔驰糖果派对攻略 作者:生物质谱实验室 时间:2009-04-21 点击:

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应用实例:

Reference
Cunzhong Yuan, Liyan Jiao ,Wantao Ying, Xiaohong Qia,et.al. The up-regulation of 14-3-3 proteins in Smad4 deficient epidermis and hair follicles at catagen. Proteomics 2008, 8, 2230–2243.

Introduction
we performed 2-D DIGE proteomic analysis of epidermal and HF keratinocytes from the Smad4 mutant and control mice during anagen and catagen to explore the differentially expressed proteins during the initiation of catagen.

Methods
1. 2-DE DIGE
Protein samples (50 mg) were labeled with 400 pmol of Nhydroxysuccinimide cyanine dyes Cy2, Cy3, or Cy5 following the experimental design shown in Fig. 1
2. Image analysis
3. In-gel digestion and MS identification
4. Database searching
5. Cluster analysis
6. Western blot and immunofluorescence
7. RNA isolation, real time (RT)-PCR, and quantitative RT-PCR (qRT-PCR)