10 of the best ELISA practices & techniques
10 of the best ELISA practices and techniques
- 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.