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Titolo:
SYSTEMATIC PEPTIDE FRAGMENTATION OF POLYVINYLIDENE DIFLUORIDE(PVDF)-IMMOBILIZED PROTEINS PRIOR TO MICROSEQUENCING
Autore:
IWAMATSU A; YOSHIDAKUBOMURA N;
Indirizzi:
KIRIN BREWERY CO LTD,CENT LABS KEY TECHNOL,KANAZAWA KU,1-13-5 FUKUURAYOKOHAMA KANAGAWA 236 JAPAN
Titolo Testata:
Journal of Biochemistry
fascicolo: 1, volume: 120, anno: 1996,
pagine: 29 - 34
SICI:
0021-924X(1996)120:1<29:SPFOPD>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
INSITU PROTEASE DIGESTION; GEL-ELECTROPHORESIS; PURIFICATION; BINDING; CDNA;
Keywords:
DEDUCIBLE AMINO ACID RESIDUES; PVDF MEMBRANE; RECOVERIES OF DIGESTS FROM PVDF; S-ALKYLATION OF PROTEIN; SEQUENTIAL IN SITU DIGESTION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
10
Recensione:
Indirizzi per estratti:
Citazione:
A. Iwamatsu e N. Yoshidakubomura, "SYSTEMATIC PEPTIDE FRAGMENTATION OF POLYVINYLIDENE DIFLUORIDE(PVDF)-IMMOBILIZED PROTEINS PRIOR TO MICROSEQUENCING", Journal of Biochemistry, 120(1), 1996, pp. 29-34

Abstract

The sequential in situ digestion of proteins immobilized on polyvinylidene difluoride (PVDF) has been systematically designed and optimized, The method consists of immobilization of the proteins on PVDF, S-carboxymethylation, and then successive in situ digestions using specificproteases, In order to obtain high yields of the peptide fragments, from which specific amino acid residues connected to the N- or C-terminal of the resulting digestion fragments can be deduced, the cleavages are performed in the following order: (1) Achromobacter protease I (API), (2) endoproteinase Asp-N, and (3) trypsin, Procedures for recovering the numerous fragments remaining on the PVDF membrane after the third digestion with trypsin are also discussed, Application of sequential in situ digestion for the acquisition of fragments suitable for sequencing from digests of large-molecular-weight proteins is also presented.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/01/21 alle ore 02:30:12