Monoclonal antibody specific for an activated RAS protein.
- 1 October 1986
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 83 (19), 7485-7489
- https://doi.org/10.1073/pnas.83.19.7485
Abstract
Activated RAS transforming genes that encode proteins (p21s) with amino acid substitutions at positions 12, 13, or 61 have been detected in 10-20% of human neoplasms. This report describes a monoclonal antibody (DWP) raised against a synthetic peptide corresponding to amino acids 5-16 of a mutated RAS gene encoding Val instead of Gly at position 12. DWP reacted in competition assays with peptides containing Val or Cys at position 12, but did not react with peptides containing Gly, Arg, Ser, Ala, Asp, or Glu at position 12. Immunoblot analysis of transformed NIH cells and human carcinoma cell lines showed that DWP reacts specifically with activated RAS proteins containing Val at position 12 and not with normal p21s or p21s activated by other amino acid substitutions at positions 12 and 61. Immunohistochemical studies showed that DWP-labeled transformed NIH cells and human carcinoma cells contained p21s with either Val or Cys at position 12 but not normal or other activated p21s. In contrast to the specificity seen with human carcinoma cell lines, analysis of formalin-fixed, primary carcinoma specimens indicated that positive immunoperoxidase staining with DWP did not necessarily correlate with immunoblot and transfection assays for the presence of activated RAS proteins. Immunohistochemical studies did show, however, that DWP preferentially binds human carcinoma cells.Keywords
This publication has 29 references indexed in Scilit:
- Biological and biochemical properties of human rasH genes mutated at codon 61Cell, 1986
- Cellular transforming genes and oncogenesisBiochimica et Biophysica Acta (BBA) - Reviews on Cancer, 1984
- Acquisition of transforming properties by alternative point mutations within c-bas/has human proto-oncogeneNature, 1983
- Altered gene products are associated with activation of cellular rasK genes in human lung and colon carcinomasCell, 1983
- A point mutation is responsible for the acquisition of transforming properties by the T24 human bladder carcinoma oncogeneNature, 1982
- Mechanism of activation of a human oncogeneNature, 1982
- Localization of the src gene product of the Harvey strain of MSV to plasma membrane of transformed cells by electron microscopic immunocytochemistryCell, 1980
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences of the United States of America, 1979
- Transformation of NIH/3T3 mouse cells by DNA of Rous sarcoma virusCell, 1979
- Antibodies to Bradykinin and Angiotensin: A Use of Carbodiimides in ImmunologyScience, 1964