High signal/absorbance in some or all wells

 

Contamination of Substrate TMB

Substrate TMB was contaminated by enzyme conjugate solution.

Always use clean pipette tips, beakers etc. in order to avoid contamination. Rinsing with water is not sufficient. Containers and pipette tips used for enzyme conjugate solution should be kept separate from the Substrate TMB.

Contamination with samples or calibrators

Assay-, Sample- or Conjugate buffer was contaminated by samples, purified antigen/protein, control or Calibrators.

Always use clean pipette tips, beakers etc in order to avoid contamination.

Washing procedure

Insufficient washing before Substrate TMB incubation.

Plate washer should be checked and served periodically. When washing manually use a laboratory wash bottle, see Manual Wash Instructions.

Incorrect dilution of enzyme conjugate solution

Too much enzyme conjugate stock solution has been added to the Enzyme Conjugate Buffer.

Dilute enzyme conjugate solution according to Directions for Use