No Signal
- Improper standard dilution
- Inccorect storage of reagents
- Incomplete washing of the wells
- Capture antibody did not bind to theplate
Solution
- Reconstitute standard according to the
- Store the reagents in the ELISA kit according to the printed instruction before using them
- Make sure wells are washed adequately by filling the wells with wash buffer and all residual antibody solutions crossed well before washing.
- Replace a new ELISA plate
High Background
- Improper washing
- Substrate was contaminated
- Non-specific binding of antibody
- Plate are not be sealing incompletely
- Incorrect incubation temperature
- Substrate exposed to light prior to use
- Contaminated wash buffer
Solution
- Increasing duration of soaking steps
- Substrate should be clean and avoid crossed contamination by using the sealer
- Replace another puritified antibody or blocking buffer
- Make sure to follow the instruction strictly
- Incubate at room temperature
- Keep substrate in a dark place
- Use a clean buffers and sterile filter.
Weak Signal
- Improper washing
- Incorrect incubation temperature
- Antibody are not enough
- Reagent are contaminated
- Pipette are not clean
Solution
- Increasing duration of soaking steps
- Incubate at room temperature
- Increase the concentration of the antibody
- Use new one
- Pipette should be clean
Poor Standard Curve
- Improper standard dilution
- Inccorect storage of reagents
- Incomplete washing of the wells
- Capture antibody did not bind to theplate
Solution
- Reconstitute standard according tothe
- Store the reagents in the ELISA kit according to theprinted instruction before using them
- Make sure wells are washed adequatelyby filling the wells with wash buffer and all residual antibody solutions crossed well before washing
- Replace a new ELISA plate