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ELISA Troubleshooting: Saturated Signal
If your ELISA signal is too high, the results of the experiment can become unusable. Saturated signal can cause wells to appear uniformly reactive, or cause the standard curve to become unusable. Before you repeat your ELISA experiment, read these troubleshooting tips to identify possible sources of your saturated signal error, and solutions to solve it.
|Substrate problems||Excessive substrate||Decrease substrate concentration|
|Incubation time too long||Reduce incubation time|
|Reduce incubation temperature|
|Substrate activation before use||Make substrates immediately before use|
|Plate error||Insufficient washing||Follow manufacturer’s washing instructions|
|Thoroughly drain plate after washes|
|Plate sealer not used or reused||Cover plate with plate sealer during incubations|
|Use fresh sealer every time|
|Plate read at incorrect wavelength||Use recommended wavelength/filter|
|Check that plate reader is set up for the type of substrate used|
|Antibody problems||Excessive antibody concentration||Reduce antibody concentration|
|Nonspecific antibody binding||Use a different formulation of coating solution|
|Use Boster antibodies guaranteed to only react with their targets|
Keywords:ELISA, Troubleshooting, Saturated Signal, O.D., Off ScaleClick for more troubleshooting tips