Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and centrifuge blotting: preparation of polypeptides for amino-terminal sequence analysis
| Autor: | Knut Sletten, Jessie Juul, Leonila F. Hermansen |
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| Rok vydání: | 1996 |
| Předmět: |
Chromatography
Time Factors Edman degradation Molecular mass Clinical Biochemistry Immunoblotting Sodium Dodecyl Sulfate Centrifugation Membranes Artificial Biochemistry Analytical Chemistry chemistry.chemical_compound Membrane chemistry Cyanogen bromide Electrophoresis Polyacrylamide Gel Polyvinyls Sodium dodecyl sulfate Southwestern blot Peptides Polyacrylamide gel electrophoresis Sequence Analysis |
| Zdroj: | Electrophoresis. 17(4) |
| ISSN: | 0173-0835 |
| Popis: | The applicability and reproducibility of a previously described (L. F. Hermansen et al., Electrophoresis 1993, 14, 1302-1306) centrifuge-blotting procedure for capturing subnanomolar amounts of protein on polyvinylidene difluoride membranes for direct Edman degradation was further investigated. Proteins with different molecular masses were centrifuge-blotted onto Immobilon CD membranes. Simultaneous blotting and desalting was achieved with an overall yield of 15-56% after 2 h centrifugation for proteins with a molecular mass of 12-30 kDa. Centrifugation of myoglobin for 6 h resulted in an overall yield of 72%. The subnanomolar amounts obtained were also sufficient to conduct cyanogen bromide cleavage in situ on proteins with blocked NH2-terminus and to generate sequence information. |
| Databáze: | OpenAIRE |
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