ELISA Plate Washing Tips

Washing of ELISA plates is critical process in maintaining consistency between resulsts, reducing intra/inter variability and producing high quality data.

At ELISA Genie we recommend the use of an ELISA plate washer where possible.

However, pipetting and washing plates by hand is also an option but may result in inconsistencies in data due to variability.

Before you begin washing your ELISA plates, ensure you have all the required tools including a multi-channel pipette, a reservoir, gloves, safety goggles, a cloth/towel/paper to damp your ELISA plates, clingfilm/plate covers.

Standard ELISA protocol and wash steps for a sandwich ELISA

6 tips to help with your ELISA plate washing protocols

  1. ELISA plate washing should be as consistent as possible from plate to plate and row to row within a plate, for this reason automated or semi-automated washing is preferred over manual washing.
  2. If using an automated option ensure that the machine is working properly and cleaned regularly. It is also important to check that the dispensing needles are working correctly, dispensing with a smooth and steady stream.
  3. When washing always wash with 300-350ul of wash buffer. Do not over fill the wells as this this may invalidate test results.
  4. Do not scrape the surface of the wells when washing as this may remove any bound antigen/antibody, resulting in inaccurate and inconsistent results.
  5. Do not allow the plate to dry out between washes as this may affect your results. Work quickly so that the time from washing the first row to the last row isnt too long and drying out is minimal.
  6. When tapping the plate out a paper towel check for evidence of colour. This may suggest that the wells have not been washed properly and there are reagents remaining in or around the wells.