In order to find the optimum concentration of my tracer, I replaced it in a commercial kit instead of kit's tracer, in the range of 5-150 ng/ml, but I was surprised to see the OD was similar and high in the whole range. (OD of kit = 1.9   ,   OD using my tracer in 5-150 ng/ml range = 1.9 – 2.1). Since the kit is designed to quantify analyte concentrations of up to 140 ng/ml, I think it’s not normal to get a maximum OD with labeled antigen as low as 5 ng/ml.

I would appreciate it very much if you could guide me if this is normal or am I doing something wrong?

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