The importance of including ELISA controls, both positive and negative, in your immunoassay helps to verify that the assay was run properly and everything is performing accurately.
Positive ELISA Controls
A positive ELISA control can be a recombinant or natural sample that you know will be detectable in the assay. Positive controls help to show that a negative sample is truly negative. The standard curve is one form of positive control and you can compare your results to the standard curve data that is provided in your product insert. R&D Systems also sells ELISA controls for the Quantikine ELISAs . Most Human Quantikine kits have a lyophilized tri-level control with expected ranges that are validated by our Quality Control and our Mouse/Rat Quantikine ELISAs include one control in the kit. These are great when running multiple plates or when you have multiple users running the assay, to verify that values are all within the expected ranges.
ELISA Spike Controls
When using complex sample matrices, it is also important to make sure that there is nothing present in the matrix that interferes in the assay. It is recommended to spike in recombinant or natural protein into your matrix and verify that the amount you spike in is what you read out. For more information, refer to Section 4f. Our Quantikine ELISAs are already validated for many complex sample types, so refer to the kit insert for details.
Negative ELISA Controls
Negative controls help to verify that you are not obtaining any false positive results or non-specific binding. Use a sample that you know will does express the protein you are measuring. If you are quantitating a cell culture supernate, a good negative control would be to test your cell culture media.
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Consistency of Control Values Over Time
Quantitation of Human IL-6 in High, Medium, and Low Controls. High (blue line), medium (red line), and low (green line) controls are assayed with every manufactured lot of the Human IL-6 Quantikine® ELISA Kit (Catalog # D6050). Controls for the Human IL-6 Quantikine® ELISA Kit fall within acceptable ranges (gray bars) and remain consistent from lot to lot.