Electroimmunoblotting of small peptides Separated on Urea-Dodecyl Sulphate (SUDS) gels application to myelin basic protein

H. Z. Sheng; R. E. Martenson; P. R. Carnegie; C. C A Bernard

doi:10.1016/0022-1759(88)90003-8

Electroimmunoblotting of small peptides Separated on Urea-Dodecyl Sulphate (SUDS) gels application to myelin basic protein

H. Z. Sheng, R. E. Martenson, P. R. Carnegie, C. C A Bernard

Research output: Contribution to journal › Article › Research › peer-review

7 Citations (Scopus)

Abstract

A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized and as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.

Original language	English
Pages (from-to)	13-22
Number of pages	10
Journal	Journal of Immunological Methods
Volume	107
Issue number	1
DOIs	https://doi.org/10.1016/0022-1759(88)90003-8
Publication status	Published - 24 Feb 1988
Externally published	Yes

Keywords

Immunoblot
Monoclonal antibody
Myelin basic protein
Peptide
Polyclonal antibody

Access to Document

10.1016/0022-1759(88)90003-8

Link to publication in Scopus

Cite this

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title = "Electroimmunoblotting of small peptides Separated on Urea-Dodecyl Sulphate (SUDS) gels application to myelin basic protein",

abstract = "A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized and as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.",

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T1 - Electroimmunoblotting of small peptides Separated on Urea-Dodecyl Sulphate (SUDS) gels application to myelin basic protein

AU - Sheng, H. Z.

AU - Martenson, R. E.

AU - Carnegie, P. R.

AU - Bernard, C. C A

PY - 1988/2/24

Y1 - 1988/2/24

N2 - A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized and as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.

AB - A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized and as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.

KW - Immunoblot

KW - Monoclonal antibody

KW - Myelin basic protein

KW - Peptide

KW - Polyclonal antibody

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