Radioimmunoassay of the APC Gene Product Using Antibodies against Its Middle and Carboxyl Regions
| Autor: | Rey Marbello, B. M. Boman, Richard F. Murphy, Thomas E. Adrian, Gautam Bhattacharya, Christine Abraham, Sándor Lovas |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Adenomatous Polyposis Coli Protein
Blotting Western Molecular Sequence Data Radioimmunoassay Biophysics Biochemistry Antibodies Epitope Antigen Antibody Specificity Humans Amino Acid Sequence Antigens Molecular Biology Peptide sequence chemistry.chemical_classification biology Proteolytic enzymes Cell Biology Molecular biology Peptide Fragments Amino acid Molecular Weight Cytoskeletal Proteins chemistry Polyclonal antibodies Colonic Neoplasms biology.protein Antibody |
| Zdroj: | Biochemical and Biophysical Research Communications. 206:909-915 |
| ISSN: | 0006-291X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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