We’re currently running a series of tests on serum samples to quantify specific protein biomarkers, and while ELISA remains our go-to method due to its sensitivity and specificity, the overwhelming number of kit options makes selection pretty confusing. Some kits differ a lot in terms of detection ranges, sample volume required, and even cross-reactivity rates. We’re trying to avoid unreliable results that could come from kits with suboptimal antibody specificity or low reproducibility. Does anyone have a structured approach to evaluating these kits before committing to one? Are there parameters beyond just the datasheet and IFU (instructions for use) that are worth considering during the decision-making process?
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When we choose ELISA kits in our lab, we usually focus on a few core factors before placing any orders. Beyond just reading through datasheets, we often look into the validation data—specifically the intra- and inter-assay CVs. If the coefficients of variation are consistently low, that's a good indication that the kit performs reliably over multiple runs and across different users. Another thing we check is whether the target biomarker was validated using real biological samples (not just standards), which gives a better sense of how the assay performs under actual conditions. Kits that only show performance with spiked controls might not behave the same way when applied to serum, plasma, or cell culture supernatants. Also, even though many kits claim cross-species reactivity, this should be interpreted carefully. Just because something works in human samples doesn’t mean you’ll get meaningful results in murine or porcine tissues. We once had an issue with this when a cytokine kit worked beautifully on one species but gave totally erratic results with another. There's actually a solid overview of ELISA kits and antibodies that outlines different kit types, detection systems, and sample requirements here: https://gentaur.co.uk. It’s not just a catalog — they explain some of the practical considerations that go into selecting the right kit, including how to match your detection system (colorimetric, chemiluminescent, or fluorescent) with your lab's available equipment.