ELISA Kit Troubleshooting: Poor Standard Curve

Though without that complicated protocols as IHC, immunoprecipitation etc, ELISA assay might generate many problems as what could occur in most of the immuno assays. This part provides troubleshooting of ELISA kit with means of cause-solution table with problems listed beforehand. Please note that the conditions described here may not pertain to every ELISA kit because performance requirements vary for individual assays. Be sure to check your package insert for specifications.

In ELISA kit, the optical density measurement often gives poor optical density (OD) readings, and Eventually lead to poor standard curve. This section is intended to recommend possible causes and relative troubleshooting for the problem of ELISA kit poor standard curve.

Possible Sources Solutions
Standard was incompletely reconstituted or was inappropriately stored Aliquot reconstituted standard and store at-80 ℃. The reconstituted standards should be aliquoted and avoid repeated freeze-thaw cycles.
Imprecise / inaccurate pipetting Check / calibrate pipettes
Incubations done at inappropriate temperature, timing or agitation Follow the general ELISA protocol
Background wells were contaminated Avoid cross contamination by using the sealer appropriately