Abstract
The transfer onto immobilizing membranes of proteins which are separated on polyacrylamide gels (popularly referred to as the Western blot) has become an important tool in protein chemistry (for a comprehensive review see [1]). Originally, transferred proteins were covalently bound onto the membrane [2, 3], but this method showed low coupling yields and was therefore replaced by the introduction of microporous nitrocellulose membranes to which proteins could be bound by noncovalent interactions [4]. This principle was later improved by using a nylon-based, positively charged membrane [5]. Although proteins immobilized in this way could be easily assayed for their antigenicity or their enzymatic or lectin-binding activity [1], the membranes used for these analyses were sensitive to the chemical treatments necessary for acid hydrolysis or for the Edman-de-gradation-based chemistry. In order to combine the simple, fast, inexpensive, high-resolution purification procedure of polyacrylamide gel electrophoresis with the recently developed technique of protein gas-phase sequencing [6], it was necessary to introduce a support which is both able to bind every kind of protein eluting from a gel, and resistant against chemicals and solvents used in Edman chemistry.
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© 1986 Springer-Verlag Berlin Heidelberg
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Vandekerckhove, J., Bauw, G., Puype, M., van Damme, J., van Montagu, M. (1986). Protein Blotting from Polyacrylamide Gels on Glass Microfiber Sheets: Acid Hydrolysis and Gas-Phase Sequencing of Glass-Fiber Immobilized Proteins. In: Wittmann-Liebold, B., Salnikow, J., Erdmann, V.A. (eds) Advanced Methods in Protein Microsequence Analysis. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-71534-1_15
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DOI: https://doi.org/10.1007/978-3-642-71534-1_15
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