Radioimmunoassay of the APC Gene Product Using Antibodies against Its Middle and Carboxyl Regions

B. M. Boman, Sándor Lovas, C. Abraham, T. E. Adrian, R. F. Murphy, R. Marbello, G. Bhattacharya

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Abstract

A radioimmunoassay (RIA) has been developed for the adenomatous polyposis coli protein (APC). High-avidity rabbit polyclonal antibodies were produced against synthetic peptides corresponding to amino acids 1865-1881 (APC-1) and to amino acids 1336-1350 (APC-2) in APC′s 2844 amino acid sequence. Both antibodies were utilized in RIA to evaluate full-length APC that is present in the insoluble particulate fraction of cell lysates. High salt extraction, often employed for coiled-coil type protein preparations, was found to be useful for extraction of APC from lysates of normal colonic epithelium. Proteolytic digestion of high salt extracts increased antibody reactivity toward both epitopes, suggesting that APC-1 and APC-2 antigenic sites are partially concealed due to APC′s involvement in multiprotein complexes. Thus, RIA using our antibodies will provide a valuable tool for APC protein purification and in studies for elucidating APC′s biologic function.

Original languageEnglish
Pages (from-to)909-915
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume206
Issue number3
DOIs
Publication statusPublished - Jan 26 1995

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All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

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