Detection of secreted and intracellular annexin II by a radioimmunoassay.
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 microgram 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 micrograms 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.
Davis
R G
RG
Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha 68198-4525, USA.
Vishwanatha
J K
JK
D000230:Adenocarcinoma / Q000737:chemistry / Q000378:metabolism; D017306:Annexin A2 / Q000032:analysis* / Q000378:metabolism*; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000737:chemistry / Q000378:metabolism; D010920:Placenta / Q000737:chemistry; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
D017306:Annexin A2