Elution of proteins from polyacrylamide eels: a simple and economic procedure

Bongertz, Vera

doi:10.1590/S0036-46651989000100009

Abstracts

A simplified methodology for the quantitative electroelution of proteins from polyacrylamide gels is described. After staining with Coomassie Brilliant Blue R 250, the identified bands are excised from the gel and the proteins eluted using a procedure developed for use in conventional tube gel electrophoresis equipment.

Descreve-se uma metodologia simplificada para a eletroeluição quantitativa de proteínas de gel de poliacrilamida. Após coloração do gel pelo Coomassie Brilliant Blue R 250, os componentes identificados são recortados e as proteínas eluidas do gel por um procedimento desenvolvido para uso em aparelho de eletroforese vertical em tubos.

Poliacrilamida; Eletroeluição; Proteínas

ENSAIOS TÉCNICOS

Vera Bongertz

Departamento de Imunologia. Fundação Oswaldo Cruz. Caixa Postal 926. CEP 20001 Rio de Janeiro, RJ, Brasil

RESUMO

Descreve-se uma metodologia simplificada para a eletroeluição quantitativa de proteínas de gel de poliacrilamida. Após coloração do gel pelo Coomassie Brilliant Blue R 250, os componentes identificados são recortados e as proteínas eluidas do gel por um procedimento desenvolvido para uso em aparelho de eletroforese vertical em tubos.

Unitermos: Poliacrilamida; Eletroeluição; Proteínas.

SUMMARY

A simplified methodology for the quantitative electroelution of proteins from polyacrylamide gels is described. After staining with Coomassie Brilliant Blue R 250, the identified bands are excised from the gel and the proteins eluted using a procedure developed for use in conventional tube gel electrophoresis equipment.

Texto completo disponível apenas em PDF.

Full text available only in PDF format.

AGRADECIMENTOS

Este trabalho foi financiado pelo Comitê de Imunologia de Leishmaniose da Organização Mundial da Saúde (Immleish - TDR-WHO) número ID 850349 e pelo CNPq (PIDE VI) projeto número 400720/85. Agradecemos a Maria Solange Correia Soeiro pela assistência técnica e a Jorge Carvalho Cruz pelos serviços fotográficos.

Recebido para publicação em 6/6/1888

1. ABOU-ZEID, C; FILLY, E.; STEELE, J. & ROCK, G. A. W. - A simple method for using antigens separated by Polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from western blots into antigen bearing particles. J. Immunol. Meth., 98: 5-10, 1987.
2. BOULARD, C. & LECROISEY, H. - Specific antisera produced by direct immunization with slices of Polyacrylamide gel containing small amounts of protein. J. Immunol. Meth., 50: 221-226, 1982.
3. GODING, J. W. & HANDMAN, E. - Electrophoretic analysis of protein antigens. In: MOREL, C. M., ed. - Genes and antigens of parasites: a laboratory manual. 2nd. ed. Rio de Janeiro, FIOCRUZ, 1984. p. 383-415.
4. HAGER, D. A. & BURGESS, R. R. - Elution of proteins from sodium dodecylsufate Polyacrylamide gels, removal of sodium dodecyl sulfate and renaturation of enzymatic activity: results with Sigma subunit of Escherichia coli RNA polymerase, Wheat Germ DNA topolsomerase and other enzymes. Anal. Biochem., 109: 76-86,1980.
5. HANAOKA, F.; SHAW, J. L. & MUELLER, G. C. - Recovery of functional proteins from sodium dodecyl sulfate - Polyacrylamide gels. Anal. Biochem., 99: 170-174, 1979.
6. LAEMMLI, U. K. - Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London), 227: 680-685, 1970.
7. TOWBIN, H.; STAEHELIN, T. & GORDON, J. - Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets. Procedure and some applications. Proc. natl. Acad. Sci. (Wash.), 76: 4350-4354, 1979.
8. TUSZYNSKI, G. P. & WARREN, L. - Removal of sodium dodecyl sulfate from proteins. Anal. Biochem., 67: 55-65, 1975.

Eluição de proteínas de gel de poliacrilamida: descrição de metodologia simples e econômica

Elution of proteins from polyacrylamide eels: a simple and economic procedure

Publication Dates

Publication in this collection
18 Feb 2011
Date of issue
Feb 1989

History

Received
06 June 1888

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ABOU-ZEID, C; FILLY, E.; STEELE, J. & ROCK, G. A. W. - A simple method for using antigens separated by Polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from western blots into antigen bearing particles. J. Immunol. Meth., 98: 5-10, 1987.

[2] 2. BOULARD, C. & LECROISEY, H. - Specific antisera produced by direct immunization with slices of Polyacrylamide gel containing small amounts of protein. J. Immunol. Meth., 50: 221-226, 1982.

[3] 3. GODING, J. W. & HANDMAN, E. - Electrophoretic analysis of protein antigens. In: MOREL, C. M., ed. - Genes and antigens of parasites: a laboratory manual. 2nd. ed. Rio de Janeiro, FIOCRUZ, 1984. p. 383-415.

[4] 4. HAGER, D. A. & BURGESS, R. R. - Elution of proteins from sodium dodecylsufate Polyacrylamide gels, removal of sodium dodecyl sulfate and renaturation of enzymatic activity: results with Sigma subunit of Escherichia coli RNA polymerase, Wheat Germ DNA topolsomerase and other enzymes. Anal. Biochem., 109: 76-86,1980.

[5] 5. HANAOKA, F.; SHAW, J. L. & MUELLER, G. C. - Recovery of functional proteins from sodium dodecyl sulfate - Polyacrylamide gels. Anal. Biochem., 99: 170-174, 1979.

[6] 6. LAEMMLI, U. K. - Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London), 227: 680-685, 1970.

[7] 7. TOWBIN, H.; STAEHELIN, T. & GORDON, J. - Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets. Procedure and some applications. Proc. natl. Acad. Sci. (Wash.), 76: 4350-4354, 1979.

[8] 8. TUSZYNSKI, G. P. & WARREN, L. - Removal of sodium dodecyl sulfate from proteins. Anal. Biochem., 67: 55-65, 1975.