10 great ELISA Tips!

10 of the best ELISA practices and techniques

Below we have put together 10 simple tips to help you with your ELISA assay. You can find a more comprehensive list in our "101 ELISA troubleshooting techniques" guide which gives a great overview of ELISA troubleshooting.

Number Technique


Make sure all reagents are at room temperature before use, unless instructed to keep them cool.


If you aren’t going to run a full plate, ensure that the remaining wells are sealed in a plate bag to avoid contamination and to prevent moisture from degrading the plate.


If your standards are not single use and you intend on using them more than once, its best to aliquot and freeze smaller volumes, this helps avoid repeated freeze-thaws.


Use a multiple channel pipette if possible, this helps with speed and results in more consistent results.


When pipetting, try and dispense liquid with the pipette tips held at an angle and not touching the bottom of the well.


Always change your pipette tips between different samples or standards to prevent contamination.


Manual plate washing can lead to higher backgrounds if possible use an automated plate washer.


Always give your wash buffer enough time to work, add a 30 second soak time between washes.


Be strict with your incubation times. Incubation times should not vary by more than 5 minutes over or under an hour incubation time.


If you are incubating multiple plates in a cold enviroment, do not stack the plates on top of each other place them individually on the shelf.