Choosing the right volume capacity for your ELISA microplate depends on several factors related to your samples and reagents:

l Sample Volume: ELISA samples typically contain 100 microliters (µL). This volume is commonly used because it strikes an appropriate balance between sensitivity and well capacity. If your sample volume is limited, consider using microplates with a 50 µL well capacity. These plates allow you to run the assay with fewer samples while retaining high sensitivity. However, make sure to check your ELISA kit's instructions for compatibility with lower volumes. If you have a large volume of samples and need to run multiple replicates or test a larger number of samples, consider using 384- or even 1536-well plates. These plates provide higher throughput but need careful optimization due to the smaller well volume (typically 16 µL).

l Reagent Volume: Standard ELISA protocols typically require the same volume of washing buffers, conjugate, and substrate solutions as the sample volume (usually 100 µL). Remember the "dead volume" of your pipettes. This is the smallest volume that remains in the pipette tip after dispensing. You must account for this when calculating the appropriate well volume to ensure that enough reagent is dispensed for the assay.

l Number of Replicates and Standards: To improve data reliability, most ELISA experiments require that samples and standards be run in duplicate or triplicate. Include this in your well volume requirements. ELISAs frequently use a standard curve to quantify the target analyte. Consider how many standard dilutions you'll need and how many wells each one will require.

l Evaporation: During the ELISA incubation steps, some liquids may evaporate. This is especially true for longer incubations at high temperatures. Choosing a suitable volume that allows for a small margin of error can help to reduce the impact of evaporation.

By carefully considering these factors, you can choose the ELISA microplate volume capacity that best suits your specific experiment and sample availability. For more information about choosing the right ELISA microplate, please click here.