AlphaLISA 5 μL vs. 20 μL Sample Volume
Increasing the volume of the sample while keeping the overall reaction volume can increase the sensitivity of the assay. This is because you are adding more molecules of analyte into the well.
Please note that increasing the volume of sample in a 50 µL reaction might also increase any potential assay interference arising from your sample or matrix. We do not recommend adjusting the volume of your sample if you are using serum samples.