ELISA kit Troubleshooting - High Background

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.

No Signal
Poor
standard
curve
Poor
Detection
Value
Non
-Specificity
High
Background
Possible Sources Solutions
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
Antibody
Antibody
ELISA Antibody
ELISA KIT + ELISA Pair set
ELISA KIT+
- ELISA Kit technology center
ELISA kit Introduction
ELISA Kit principle
ELISA kit protocol
ELISA kit troubleshooting
ELISA kit Troubleshooting-No signal
ELISA kit Troubleshooting - Poor Standard Curve
ELISA kit Troubleshooting - Poor detection value
ELISA kit Troubleshooting - High Background
ELISA kit Troubleshooting - Non-specificity
ELISA kit FAQ
ELISA kit Tips
ELISA kit storage
ELISA kit material provided
ELISA Kit precaution and safety instructions
ELISA Kit Terms