Download Analisi Due by Gianni Gilardi PDF

By Gianni Gilardi

ISBN-10: 8838607273

ISBN-13: 9788838607271

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Electrophoresis 16: 438-443 43 Aebersold RH, Leavitt J, Saavedra RA, Hood LE, Kent SBH (1987) Internal amino acid sequence analysis of proteins separated by one- and two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose. Proc Natl Acad Sci USA 84: 69706974 44 Hess D, Aebersold R (1994) Internal sequence analysis of proteins separated by polyacrylamide gel electrophoresis. Methods: a companion to methods in enzymology 6: 227-238 45 Matsudaira P J (1987) Sequence from picomole quantitites of proteins electroblotted onto polyvinylidene difluoride membranes.

CNBr cleavage was demonstrated, and with some manual intervention the elution of peptides from digested protein gel slices onto an RP-HPLC column [13, 15] was performed. The digestion column was thus coupled in front of an RP-HPLC column, allowing the on-line separation of the peptide mixture. g. no blank sample can be processed as a control) as well as direct transfer to techniques such as off-line MALDI-MS and NanoES-MS. Processing of multiple samples, using a principle similar to the above, was introduced by Perseptive Biosystems (now Perkin-Elmer) [16] and BioMolecularTechnologies (now with Amersham Pharmacia Biotech) [17]; however, the set-ups are so far only applicable to proteins in solution.

Protein and peptide solutions having high salt concentrations can be infused into a microdialysis tube directly interfaced with a NanoES source. An increased signal-to-noise ratio of a factor of more than 40 is obtainable with this desalting step. However, the method is time-consuming, and sample losses sometimes occur. Another method to reduce disturbants and increase mass sensitivity is on-line coupling ofLC (liquid chromatography) to MS. But as NanoES has no need for pumps and valves, such an on-line coupling would constitute a critical set-up.

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