Automated Wash Method

    1. Use a program that will aspirate and dispense 300-400 μL wash buffer.
    2. Ensure that the plate washer has a program that will not scratch the bottom of the microplate and will prevent plate drying. This is critical to prevent damage to the capture antibody arrays. The simplest method to avoid plate drying is to leave a small, uniform amount (1-3 μL) of wash in the well after the final aspiration, and add the next reagent to the plate as quickly as possible.
    3. Leaving a uniform amount of wash in the wells can be accomplished with an automated washer by slightly increasing the aspiration height of the washer head.
      For example:
Process Distance Steps on Biotek ELx-405
Aspiration Height 3.810 mm 30
Aspiration Position 1.28 mm from center -28
Dispense Height 15.24 mm 120
No soak or shake cycles are needed

*Apiration positions can vary between individual instruments and should be adjusted

    1. The dispense rate of any automatic washer should be low to avoid washing bound material off the bottom of the plate. A dispense rate of 3, on a scale of 1-10 where 1 is the lowest, is recommended.
    2. Connect the prepared wash buffer to your automatic plate washer.
    3. Run 1-2 priming cycles to make sure that the wash buffer is running through the plate washer. When the buffer has run through the machine, the waste will be foamy.
    4. To ensure that all pins are functioning, in a spare microtiter plate, dispense 100μL wash buffer and ensure that all pins dispense uniformly, then aspirate and ensure that all pins aspirate uniformly.
    5. Prime the plate washer one time before the first wash step. When running the assay, perform the wash 3 or 6 times according to the protocol.