A preparative method for sequencing proteins and peptides:In situ gel staining with subsequent passive elution onto polyvinylidine difluoride membranes
| Autor: | Robert S. Warlow, Poornima Rajasekariah, Ronald S. Walls, Nesrin Oszarac, Andrew A. Gooley |
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| Rok vydání: | 1995 |
| Předmět: |
Molecular Sequence Data
Clinical Biochemistry Peptide Biochemistry Analytical Chemistry Humans Amino Acid Sequence Peptide sequence Gel electrophoresis chemistry.chemical_classification Chromatography Staining and Labeling Elution Difluoride Proteins Membranes Artificial Amino acid Electrophoresis Membrane chemistry Leukocytes Mononuclear Receptors Histamine Electrophoresis Polyacrylamide Gel Polyvinyls Peptides Gels Sequence Analysis |
| Zdroj: | Electrophoresis. 16:84-91 |
| ISSN: | 1522-2683 0173-0835 |
| Popis: | A preparative method for obtaining both N-terminal and internal peptide amino acid sequences from purified proteins is reported. The methodology reliably yields high fidelity signal from between 14 to 30 residues per purified protein or peptide, with low backgrounds on amino acid analysis. The procedure relies on the use of in situ staining of proteins during preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the utilisation of microconcentrators to repeatedly concentrate small amounts of proteins onto a small polyvinylidene difluoride (PVDF) disc until sufficient amounts have been adsorbed so as to give a strong sequencing signal. The protein elution and subsequent adsorption can be monitored visually with a dye and the final product, a PVDF disc with the adsorbed protein or peptide, can be directly inserted into the automated amino acid sequencer. |
| Databáze: | OpenAIRE |
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