Sequencing of proteins from two-dimensional gels by using in situ digestion and transfer of peptides to polyvinylidene difluoride membranes: application to proteins associated with sensitization in Aplysia.

@article{Kennedy1988SequencingOP,
  title={Sequencing of proteins from two-dimensional gels by using in situ digestion and transfer of peptides to polyvinylidene difluoride membranes: application to proteins associated with sensitization in Aplysia.},
  author={Timothy E Kennedy and Mary Ann Gawinowicz and Ari Barzilai and Eric R Kandel and J David Sweatt},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={1988},
  volume={85 18},
  pages={7008-12}
}
We have developed a method for obtaining partial internal amino acid sequence data from proteins isolated directly from preparative two-dimensional polyacrylamide gels. Proteins from a crude cell homogenate are separated using preparative two-dimensional polyacrylamide gel electrophoresis. Then, the gel is stained with Coomassie blue and the protein spots of interest are cut out. The in situ protein is digested with Staphylococcus aureus V8 protease in a second polyacrylamide gel and the… CONTINUE READING
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