TY - JOUR
T1 - Detection of secreted and intracellular annexin II by a radioimmunoassay
AU - Davis, Randall G.
AU - Vishwanatha, Jamboor K.
N1 - Funding Information:
The authorst hankfully acknowledgeT ony Hunter for the monoclonal antibody (D1/274.5), Anthony Floreani for providing us BALF, William Chaney for advice on radioimmunoassaya nd Yungping Chiang for providing us with purified annexin II. This research was supported in part by grants from the National Institutes of Health (GM46459) and Nebraska Cancer and Smoking Diseases Program (95-71).
PY - 1995/12/15
Y1 - 1995/12/15
N2 - 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.
AB - 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.
KW - Annexin II
KW - Bronchoalveolar lavage fluid
KW - Cell extract
KW - Pancreatic cancer
KW - Radioimmunoassay
UR - http://www.scopus.com/inward/record.url?scp=0029584858&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(95)00207-3
DO - 10.1016/0022-1759(95)00207-3
M3 - Article
C2 - 8551043
AN - SCOPUS:0029584858
SN - 0022-1759
VL - 188
SP - 91
EP - 95
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1
ER -