I’ve detected differences in frost resistance among pine seedlings cultivated under different N concentration, and I believe this response is related to the relative ability of each phenotype to synthetize CORs as a function of N availability during hardening.
First of all what technique you are using to isolate and separate your proteins? If you have used 2D Electrophoresis you can do the analysis through Image master project software or Melanie software. Beside these you can use ImageJ software for band analysis as well.
What about a proteomic differential display analysis: i.e. two-dimensional gel electrophoresis (2-DE) and subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS)?
There are many ways by which you can analyze the cold modulated proteins. The first and most important step is to extract the proteins from the non acclimated and cold acclimated Pinus seedlings. After extraction, the proteins could be resolved on SDS-PAGE or two dimensional gel electrophoresis to analyze the cold responsive proteins which can be eluted from gel and can be identified by LC-MS/MS. In addition, if you want to go for high-throughput analysis of cold modulated proteins, go for gel free analysis. iTRAQ would be a best option in this case.
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