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.

Make sure all reagents are brought to room temperature

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

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.

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.

Multichannel pipettes

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

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.

change pipettes between different samples

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.

plate washer is used as manual plate washing

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

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.

Pay close attention to the incubation times

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.

plate washer is used as manual plate washing

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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