The quickest way to determine the reason for your microplate assay not performing is to carry out the manual spectrophotometer assay 1st using a blank and the supplied kit standard. This will determine if the assay is working correctly or not.
If the assay has not worked in this manual assay please send labelled results in the MegaCalc format applicable to the kit in question which can be downloaded from the relevant kit product page.
If the kit has worked in manual format then one can assume it is the downscale to microplate volumes that is the issue, and the following tips may help:
- Some plate shakers do not supply enough agitation of the wells. We recommend to manually, and very carefully mix the wells with a clean pipette tip or by aspiration, ensuring not to splash or cross contaminate.
- Ensure pipettes are calibrated.
- Ensure the administration of the small volumes involved in microplate procedures is executed correctly and accurately.
- Ensure the standards used fall within the linear range of the kits capability. The linear range for the microplate assay can be found as shown:
