I performed an immunoassay in a microchannel (0.4 mm x 6 mm x 60 mm), with 50 ul and 25 ul sample volumes. I am getting the same reading for both the volumes across numerous tries. Is this regular, or do you think there might be something wrong?
Spiked sample was injected at 25 ul/min flow rate, followed by primary antibody bound ligand at the same flow rate. This was followed by washing buffer. Finally the reading was found to be in the same range, which was counter-intuitive. Could someone tell me why could this be happening?
Is it possible you are saturating the assay with too much analyte (perhaps both 50 and 25µl of sample contains analyte above the working range of this assay)? This is assuming you are working with a quantitative immunoassay, not simply a qualitative test. Try diluting your sample further before analysis to see if this brings it into the assay's working concentration range.
Thanks for the suggestion...though I repeated the assay today and got the right readings. There was some error in taking readings.
Dear Rohan ,
Rightly stated by Kevin M C. You should use different dilution of the sample and compare the results, you will get the clue.
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