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Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels

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Abstract
Coomassie blue-stained, heat-dried, and computer-imaged two-dimensional gels used to develop comprehensive human protein data bases served as the protein source to generate partial amino acid sequences. The protein spots were collected from multiple gels, rehydrated, concentrated by stacking into a new gel, electroblotted onto inert membranes, and in situ-digested with trypsin. Peptides eluting from the membranes were separated by HPLC and sequenced. Using this procedure, it was possible to generate partial sequences from 13 human proteins recorded in the amnion cell protein data base. Eight of these sequences matched those of proteins stored in data bases, demonstrating that a systematic analysis of proteins by computerized two-dimensional gel electrophoresis can be directly linked to protein microsequencing methods. The latter technique offers a unique opportunity to link information contained in protein data bases derived from the analysis of two-dimensional gels with forthcoming DNA sequence data on the human genome.

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Chicago
Bauw, Guy, Jozef Van Damme, Magda Puype, Joël Vandekerckhove, B Gesser, GP Ratz, JB Lauridsen, and JE Celis. 1989. “Protein-electroblotting and -microsequencing Strategies in Generating Protein Data Bases from Two-dimensional Gels.” Proceedings of the National Academy of Sciences of the United States of America 86 (20): 7701–7705.
APA
Bauw, G., Van Damme, J., Puype, M., Vandekerckhove, J., Gesser, B., Ratz, G., Lauridsen, J., et al. (1989). Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 86(20), 7701–7705.
Vancouver
1.
Bauw G, Van Damme J, Puype M, Vandekerckhove J, Gesser B, Ratz G, et al. Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. 1989;86(20):7701–5.
MLA
Bauw, Guy, Jozef Van Damme, Magda Puype, et al. “Protein-electroblotting and -microsequencing Strategies in Generating Protein Data Bases from Two-dimensional Gels.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 86.20 (1989): 7701–7705. Print.
@article{1909756,
  abstract     = {Coomassie blue-stained, heat-dried, and computer-imaged two-dimensional gels used to develop comprehensive human protein data bases served as the protein source to generate partial amino acid sequences. The protein spots were collected from multiple gels, rehydrated, concentrated by stacking into a new gel, electroblotted onto inert membranes, and in situ-digested with trypsin. Peptides eluting from the membranes were separated by HPLC and sequenced. Using this procedure, it was possible to generate partial sequences from 13 human proteins recorded in the amnion cell protein data base. Eight of these sequences matched those of proteins stored in data bases, demonstrating that a systematic analysis of proteins by computerized two-dimensional gel electrophoresis can be directly linked to protein microsequencing methods. The latter technique offers a unique opportunity to link information contained in protein data bases derived from the analysis of two-dimensional gels with forthcoming DNA sequence data on the human genome.},
  author       = {Bauw, Guy and Van Damme, Jozef and Puype, Magda and Vandekerckhove, Jo{\"e}l and Gesser, B and Ratz, GP and Lauridsen, JB and Celis, JE},
  issn         = {0027-8424},
  journal      = {PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA},
  language     = {eng},
  number       = {20},
  pages        = {7701--7705},
  title        = {Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels},
  url          = {http://dx.doi.org/10.1073/pnas.86.20.7701},
  volume       = {86},
  year         = {1989},
}

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