Detection of secreted and intracellular annexin II by a radioimmunoassay

Randall G. Davis, Jamboor K. Vishwanatha

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)91-95
Number of pages5
JournalJournal of Immunological Methods
Volume188
Issue number1
DOIs
StatePublished - 15 Dec 1995

Keywords

  • Annexin II
  • Bronchoalveolar lavage fluid
  • Cell extract
  • Pancreatic cancer
  • Radioimmunoassay

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