I'm working in banana proteomics. Normally I am getting protein pellet in very low quantity. After getting the pellet I suspend the pellet in 1x SDS sample buffer. Then I boil the sample for 5 mins at 100 degree C and load in the gel but once I boiled the sample it means it is not at all possible to use the sample for 2D. So is there any common buffer available for both normal SDS PAGE and 2D?
See this ms
http://www.bio.puc.cl/caseb/pdf/prog7/jpy_07-253.pdf
and
http://www.intechopen.com/books/gel-electrophoresis-advanced-techniques/proteomics-in-seaweeds-ecological-interpretations
You have to be careful, SDS-Sample Buffer is not usable for 2D-gels. SDS-Buffer is used for 1D-gels to solubilize the proteins and in combination with the heat to unfold the proteins. In 2D buffer you have urea and thiourea, but urea will degradate when it is heated over 37°C.
I dissolved the protein pallet in deionized water for SDS PAGE and in rehydration buffer for 2D. So far no common buffer is known. You should prepare pallet in duplicate
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