Hello everyone,
I have a question concerning IL-10 secretion by murine M2 macrophages. I am currently doing an ELISA test to evaluate the secretion, but first I would like to know what is the secretion range for about 100 000 cells ? This information would be very useful in order to know if my datea will fit with the standard curve (references with standard protein).
Thanks a lot,
Maylis
It is hard to predict IL-10 levels, or the levels of any cytokine for that matter. It would depend on how you polarized your M2 macrophages, what you are stimulating them with, how long you are stimulating them for, how may cells you have per well, in what volume of media and so on. Even if you manage to replicate your experimental setup, ELISA values could differ from one assay to the next, due to inherent variability when working with biological systems (hopefully not by much). It is always best to do a preliminary run with a 2-3 fold serial dilution and then adjust the dilutions based on the data.
I completely understand the frustration of trying to get an ELISA to work on the first try. Unfortunately, this will all depend on 1) the conditions of your stim (e.g. reagents used, concentration of stimulants, how long of a stim, etc.), 2) any intervention (e.g. siRNA knockdown, knockout, drug treatment, etc.), 3) volume of the media, and 4) number of cells.
I would recommend possibly searching in the literature for someone who maybe performed a similar experiment as a starting point. But unfortunately your best bet will be to just testing your conditions with a serial dilution of a few of your experimental samples. I would highly recommend doing this if you're looking at an IL-10 ELISA as an important read-out for your experiments, especially if you're going to be doing this same ELISA several times with different conditions.
Dear I will advise you to follow the manufacturer's instructions strictly and establish the range yourself based on the cells you are working with.
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