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Protein Blotting and Detection pp 307–312Cite as

Intermittent Microwave Irradiation Facilitates Antigen-Antibody Reaction in Western Blot Analysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 536))

Summary

We established a shortened protocol for western blot analysis using intermittent microwave irradiation. With this method, the procedure is completed within 1 h after applying the primary antibody, and thus greatly saves time. This procedure appears to be applicable to any antibody based on our experience of several years.

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References

  1. Li, W., Murai, Y., Okada, E., Matsui, K., Hayashi, S., Horie, M., et al (2002) Modified and simplified western blotting protocol: use of intermittent microwave irradiation (IMWI) and 5% skim milk to improve binding specificity. Pathol Int. 52, 234–238.

    Article  PubMed  Google Scholar 

  2. Leong, A. and Milios, J. (1986) Rapid immunoperoxidase staining of lymphocyte antigens using microwave irradiation. J Pathol. 148, 183–187.

    Article  PubMed  CAS  Google Scholar 

  3. Shi, S., Cote, R., and Taylor, C. (1997) Antigen retrieval immunohistochemistry: past, present, and future. J Histochem Cytochem. 45, 327–343.

    Article  PubMed  CAS  Google Scholar 

  4. Kitayama, Y., Igarashi, H., and Sugimura, H. (2000) Initial intermittent microwave irradiation for fluorescence in situ hybridization analysis in paraffin-embedded tissue sections of gastrointestinal neoplasia. Lab Invest. 80, 779–781.

    Article  PubMed  CAS  Google Scholar 

  5. Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76, 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  6. Burnette, W. (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 112, 195–203.

    Article  PubMed  CAS  Google Scholar 

  7. Harlow, E. and Lane, D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

    Google Scholar 

  8. Toyokuni, S., Uchida, K., Okamoto, K., Hattori-Nakakuki, Y., Hiai, H., and Stadtman, E.R. (1994) Formation of 4-hydroxy-2-nonenal-modified proteins in the renal proximal tubules of rats treated with a renal carcinogen, ferric nitrilotriacetate. Proc Natl Acad Sci USA 91, 2616–2620.

    Article  PubMed  CAS  Google Scholar 

  9. Toyokuni, S., Kawaguchi, W., Akatsuka, S., Hiroyasu, M., and Hiai, H. (2003) Intermittent microwave irradiation facilitates antigen–antibody reaction in western blot analysis. Pathol Int. 53, 259–261.

    Article  PubMed  Google Scholar 

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Acknowledgements

This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan, a Grant-in-Aid for Cancer Research from the Ministry of Health, Labour and Welfare of Japan, and the Eiko Yasuhara Memorial Fund.

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Authors and Affiliations

  1. Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan

    Yu-Ting Liu & Shinya Toyokuni

Authors
  1. Yu-Ting Liu
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  2. Shinya Toyokuni
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Corresponding author

Correspondence to Shinya Toyokuni .

Editor information

Editors and Affiliations

  1. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    Biji T. Kurien

  2. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    R. Hal Scofield

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Liu, YT., Toyokuni, S. (2009). Intermittent Microwave Irradiation Facilitates Antigen-Antibody Reaction in Western Blot Analysis. In: Kurien, B., Scofield, R. (eds) Protein Blotting and Detection. Methods in Molecular Biology, vol 536. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-542-8_33

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  • DOI: https://doi.org/10.1007/978-1-59745-542-8_33

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-73-2

  • Online ISBN: 978-1-59745-542-8

  • eBook Packages: Springer Protocols

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