ELISA troubleshooting

Hundreds of variables influence ELISA data. Below is a collection of general assay running tips and troubleshooting strategies that you may find helpful when trying to resolve ELISA issues. For Q-View Data Analysis tips, please see http://www.quansysbio.com/data-analysis-q-view.

When running a Q-Plex™ Array:
Do’s
  • DO be exact when calibrating shaker speed, being off by even 100 RPM can affect results
  • DO dilute all sample types at least 1:2 (50%) with sample diluent (except cell culture media, which can be tested NEAT)
  • DO load all standards and samples into the microplate within 10 minutes of each other
  • DO be exact with incubation times, particularly the SHRP incubation
  • DO be exact when mixing Substrate A and B, being off by even 50 µL can affect results, and mix thoroughly
Don’ts
  • DON’T allow the plate to dry out between steps, particularly between washing SHRP and adding substrate
  • DON’T allow the SHRP, substrate, or IR dye to be exposed to UV light, as this may degrade it
  • DON’T analyze from a color or jpeg image; save grayscale images using a lossless image file type such as Tiff or RAW
Q-Plex Experimental Troubleshooting
Image Troubleshooting

We take great care to ensure that our products are suitable for use with all valid samples. If you have any questions about troubleshooting, or about our products or services, please contact us at 888-QUANSYS (782-6797) or info@quansysbio.com