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Twin-Site ELISAs for fos and myc Oncoproteins Using the AMPAK System

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Book cover Immunochemical Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 10))

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Abstract

Twin-site ELISA is a simple technique for quantitation of specific proteins in cell or tissue extracts. The application of this method to fos and myc oncoproteins is described. There are two basic procedures:

  1. 1.

    Extraction of fos and myc proteins from biological material in a form suitable for immunoassay.

  2. 2.

    Determination of the amount of fos or myc protein in the extract by ELISA using specific antibodies, one of which is conjugated to alkaline phosphatase (AP) and is detected by the AMPAK™ amplifier system.

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References

  1. Alitalo, K., Koskinein, T. P., Saksela, K, Sistonen, L., and Winqvist, W. (1987) Myc oncogenes: Activation and amplification. Biochimica et Biophysica Ada 907, 1–32.

    CAS  Google Scholar 

  2. Distel, R. J., Ro, R.-S., Rosen, B. S., Groves, D. L., and Spiegelman, B. M. (1987) Nucleoprotein complexes that regulate gene expression in adipocyte differentiation: Direct participation of c-fos Cell 49, 835–844.

    Article  CAS  PubMed  Google Scholar 

  3. Sassone-Corsi, P., Sisson, J. C., and Verma, I. M. (1988) Transcriptional autoregulation of the proto-oncogene fos. Nature 334, 314–319.

    Article  CAS  PubMed  Google Scholar 

  4. Heikkila, R., Schwab, G., Wikstrom, E., Loke, S. L., Pluznik, D. H., Watt, R., and Neckers, L. M. (1987) A c-myc antisense oligodeoxynucleotide inhibits entry into S phase but not progress from G0 to G2. Nature 328, 445–449.

    Article  CAS  PubMed  Google Scholar 

  5. Moore, J. P., Hancock, D. C., Iittlewood, T. D., and Evan, G. I. (1987) A sensitive and quantitative enzyme-linked immunosorbence assay for the c-myc and N-myc oncoproteins. Oncogene Res. 2, 65–80.

    CAS  PubMed  Google Scholar 

  6. Moore, J. P., Iittlewood, T. D., Hancock, D. C., and Evan, G. I. (1988) A sensitive enzyme-linked immunosorbence assay for c-fos and v-fos oncoproteins. Biochimica et Biophysica Acta 964, 60–67.

    Article  Google Scholar 

  7. Bates, D. L. (1987) Enzyme amplification in diagnostics. Trends Bwkchnol 5, 204–209.

    CAS  Google Scholar 

  8. Johannsson, A. and Bates, D. L. (1988) Amplification by second enzymes, in ELISA and Other Solid Phase Immunoassays (Kemeny, D. M. and Challacombe, S. J., eds.), John Wiley, NY, pp. 88–106.

    Google Scholar 

  9. Erisman, M. D., Scott J. K, Watt, R. A., and Astrin, S. M. (1988) The c-myc protein is constitutively expressed at elevated levels in colorectal carcinoma cell lines. Oncogene 2, 367–378.

    CAS  PubMed  Google Scholar 

  10. Mehmet, H., Sinnett-Smith, J. W., Moore, J. P., Evan, G. I., and Rosengurt, G. (1988) Differential induction of c-fos and c-myc by cyclic AMP in Swiss 3T3 cells: Significance for the mitogenic response. Oncogene Res. 3, 281–286.

    CAS  PubMed  Google Scholar 

  11. Mehmet, H., Moore, J. P., Sinnett-Smith, J. W., Evan, G. I., and Rosengurt, G. (1989) Dissociation of c-fos induction from protein kinase C-independent mitogenesis in Swiss 3T3 cells. Oncogene Res. 4, 215–227.

    CAS  PubMed  Google Scholar 

  12. Field J. K., Spandidos, D. A., Stell, P. M., Vaughan, E. D., Evan, G. I., and Moore, J. P. (1989) Elevated expression of the c-myc oncoprotein correlates with poor prognosis in head and neck squamous cell carcinoma. Oncogene Res. 4, 1463–1468.

    CAS  Google Scholar 

  13. Spandidos, D. A., Field J. K., Agnatis, N J., Evan, G. I., and Moore, J. P. (1989) High levels of c-myc protein in human breast tumours determined by a sensitive ELISA technique. Antxcancer Res. 9, 821–826.

    CAS  Google Scholar 

  14. Ishikawa, E., Imagawa, M., Hashida, S., Yoshitake, S., Hamaguchi, Y., and Ueno, T. (1983) Enzyme-labelling of antibodies and their fragments for enzyme immunoassay and histochemical staining. J. Immunoassay 4, 209–327.

    Article  CAS  PubMed  Google Scholar 

  15. Watt, R., Shatzman, A. R., and Rosenberg, M. (1985) Expression and characterization of the human c-myc DNA binding protein. Mol. Cell. Biol. 5, 448–456.

    CAS  PubMed Central  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Aaron Diamond AIDS Research Center, New York, New York

    John P. Moore

  2. Novo Nordisk Diagnostics Ltd., Cambridge, UK

    David L. Bates

Authors
  1. John P. Moore
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  2. David L. Bates
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Editor information

Editors and Affiliations

  1. Medical Research Council Laboratories, Carshalton, Surrey, UK

    Margaret M. Manson

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© 1992 The Humana Press, Inc., Totowa, NJ

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Moore, J.P., Bates, D.L. (1992). Twin-Site ELISAs for fos and myc Oncoproteins Using the AMPAK System. In: Manson, M.M. (eds) Immunochemical Protocols. Methods in Molecular Biology, vol 10. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-204-3:273

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  • DOI: https://doi.org/10.1385/0-89603-204-3:273

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-204-0

  • Online ISBN: 978-1-59259-497-9

  • eBook Packages: Springer Protocols

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