10 Best ELISA Practices and Techniques

While R&D Systems builds Quantikine ELISA kits to be robust in the hands of even inexperienced users, there are several tips and tricks that can help even the experienced user get the most from their assay.

ELISA Best Practices: Make sure all reagents are brought to the room temperature.

1) Make sure all reagents are brought to room temperature before using (unless instructed to keep them cool)

ELISA Best Practices: Prevent moisture from degrading the plate.

2) If you are not going to run the entire plate, ensure that the remaining strips are sealed in the plate bag with the desiccant to prevent moisture from degrading the plate.

ELISA Best Practices: Aliquot the remaining standard into smaller volumes and freeze.

3) For standards that are not single use, it is best to aliquot the remaining standard into smaller volumes and freeze. This allows you to avoid repeated freeze-thaws.

ELISA Best Practices: Multichannel pipettes speed the ability to plate your standard and samples.

4) Multichannel pipettes speed the ability to plate your standard and samples and lead to more consistent reseults.

ELISA Best Practices: Dispense liquid with the pipette tips held at an angle.

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

ELISA Best Practices: Use a plate washer.

6) While it is not necessary to change your pipette tips between each replicate, it is recommended that you change them between different samples or standards to prevent contamination.

ELISA Best Practices: Use a plate washer.

7) It is highly recommended that a plate washer is used as manual plate washing can lead to higher backgrounds.

ELISA Best Practices: Give the wash buffer time to work by adding a 30 second soak time in between washes.

8) When washing plates, either manually or with a plate washer, be sure to give the wash buffer time to work by adding a 30 second soak time in between washes.

ELISA Best Practices: Incubation time should not vary by more than +/-5 minutes per hour of incubation time.

9) Pay close attention to the incubation times. As a general guide the incubation time should not vary by more than +/-5 minutes per hour of incubation time.

ELISA Best Practices: Do not stack the plates on top of each to maintain cold environment.

10) If the assay calls for incubation in a cold environment, at 2-8°C, and you are running multiple assays, do not stack the plates on top of each other instead placing them individually on the shelf.

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