Detection of total IL-5 in cell supernatant

Jeff Bechard jbech at
Fri Jan 2 15:56:18 EST 1998

I am currently using a pair of matched anti mouse IL-5 antibodies in a
sandwich style ELISA to detect IL-5 in the cell supernatants of MC/9
cells. I am concerned that I may be measuring only a small percentage of
the total IL-5 present in the samples due to interference from soluble
IL-5 receptors and/or serum protein.
I have encountered this problem in the past when measuring TNF, but was
able to solve it by using a polyclonal based competitive EIA. To my
surprise the typical sandwich ELISA measured only 10% of the TNF
detected by the competitive EIA.
Unfortunately, I have been unable to source a polyclonal based
competitive EIA specific for mouse IL-5 and am hesitant to purchase the
antibody and produce my own. Does anyone have any recommendations on how
to quantitatively measure total IL-5 in cell supernatant without
interference from soluble receptors or other proteins that may bind?

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