Possible cause and recommended solutions:

  • Pipetting errors in preparation of standards > Avoid pipetting small volumes (<5 µL) and be careful not to splash from well to well
  • Air bubbles formed in well(s) > Use reverse pipetting technique
  • Standard stock is at incorrect concentration > Always refer to dilutions described in Assay preparation
  • Improperly thawed reagents > Thaw all components completely and mix well before use
  • Use of improperly stored reagents > Store the components appropriately and use fresh components from the standard curve
  • Incorrect incubation times or temperatures > Refer to Assay protocol