Detection of secreted and intracellular annexin II by a radioimmunoassay

• Published in: Journal of immunological methods Vol. 188; no. 1; pp. 91 - 95
• Main Authors: Davis, Randall G., Vishwanatha, Jamboor K.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.12.1995; Elsevier
• Subjects: Adenocarcinoma - chemistry; Adenocarcinoma - metabolism; Annexin A2 - analysis; Annexin A2 - metabolism; Annexin II; Biological and medical sciences; Bronchoalveolar lavage fluid; Bronchoalveolar Lavage Fluid - chemistry; Cell extract; Fundamental and applied biological sciences. Psychology; Fundamental immunology; HeLa Cells; Humans; Intracellular Fluid - chemistry; Intracellular Fluid - metabolism; Molecular immunology; Pancreatic cancer; Pancreatic Neoplasms - chemistry; Pancreatic Neoplasms - metabolism; Placenta - chemistry; Radioimmunoassay; Techniques; Tumor Cells, Cultured; Cell extract; Bronchoalveolar lavage fluid; Annexin II; Radioimmunoassay; Pancreatic cancer; Performance evaluation; Human; Bronchoalveolar lavage; Secretory protein; Intracellular; Detection; Hela cell line
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(95)00207-3

A simple radioimmunoassay for detection of secreted and intracellular annexin II in human cells is presented. Annexin II is a multifunctional protein in human cells and may have a role in several types of cancers. No enzymatic activity has been associated with the protein, thus making its detection difficult. Using purified annexin II from human placenta, we have developed a sensitive radioimmunoassay protocol. A linear response was observed up to a concentration of 0.5 μg purified protein in the assay. Using this radioimmunoassay protocol, annexin II can be detected in undiluted clinical human samples such as bronchoalveolar lavages and various tissue extracts. We demonstrate the applicability of this technique to measure intracellular annexin II in extracts of a human adenocarcinoma cell line (HeLa) and secreted annexin II from bronchoalveolar lavage fluid from a human patient. Using HeLa cell extracts and BAL, we observed a linear response with up to 10 μg total protein in the assay. We further demonstrate the applicability of this technique to measure differences in intracellular and secreted annexin II in the human pancreatic adenocarcinoma cell lines CD-11, CD-18 and Capan-2. While CD-11 and CD-18 do not secrete annexin II, the cell line Capan-2 secretes high-levels of the protein.