Summary
Proteins, which are characteristic for a specific state of differentiation, the transformed phenotype or pathological conditions of human cells and tissues were identified by computer analyzed two-dimensional gel electrophoresis. Sequenceable amounts of protein were collected from multiple gels with a gel-concentration device, enabling the elution and concentration of more than twenty protein spots, suspended in 1 ml of sample buffer. The eluted protein was concentrated in a new gel in a very small spot and then electroblotted onto polybase-coated glass-fiber or polyvinylidene-difluoride membranes and in situ digested. The released peptides were separated by micro-bore or narrow-bore reversed phase HPLC and immediately collected on polyethylenimine-coated glass-fiber discs for sequencing. These variations of previously developed methods allowed us to work at higher sensitivity. The procedure is currently being used to try out a systematic analysis of human proteins recovered from two-dimensional gels.
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Rasmussen, H.H. et al. (1991). Protein-Electroblotting and Microsequencing in Establishing Integrated Human Protein Databases. In: Jörnvall, H., Höög, JO., Gustavsson, AM. (eds) Methods in Protein Sequence Analysis. Advances in Life Sciences. Birkhäuser, Basel. https://doi.org/10.1007/978-3-0348-5678-2_9
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DOI: https://doi.org/10.1007/978-3-0348-5678-2_9
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