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.
ELISA high background, often means excessive color development or high optical density (OD)readings. However, sometimes, there's a high OD reading yet the color development is not dark. Here are possible causes and recommended troubleshooting for the problem of high background in ELISA. Mostly, sufficient washing or contamination avoidance should be paid more attention.
|Insufficient washes||Use multichannel pipettes without touching the reagents on the plate.|
|Insufficient washes||Increase cycles of washes and soaking time between washes|
|Color Reagent should be clear and colorless prior to addition to wells||Color Reagent should be clear and colorless prior to addition to wells|
|Use clean tubes and pipettes tips||Use clean plates, tubes and pipettes tips|