ELISA FAQ

Frequently asked questions for ELISA

  1. Are these two antibodies suitable for use together in sandwich ELISA?
  2. Is there a suitable protein standard available to use in ELISA?
  3. What dilution range should I use for the samples?

1. Are these two antibodies suitable for use together in sandwich ELISA?

Sandwich ELISA procedures can be difficult to optimize and tested match pair antibodies should be used. This ensures the antibodies are detecting different epitopes on the target protein so they do not interfere with the other antibody binding. Therefore, we are unable to guarantee our antibodies in sandwich ELISA unless they have been specifically tested for sandwich ELISA. Please review antibody datasheets for information on tested applications and details of any available matched pairs (sELISA).

2. Is there a suitable protein standard available to use in ELISA?

If there is an immunogen peptide available for use as an ELISA standard, this will be indicated on the datasheet. If you are using an ELISA kit, these often include a protein standard. We recommend checking the individual datasheet for further information.

Otherwise, you will need to search for either a full length purified endogenous or recombinant protein from the species against which the antibody was raised. Or you can choose a shorter peptide from this species, in which case you will need to ensure this contains the immunogen sequence.

3. What dilution range should I use for the samples?

This will require individual optimization for your individual sample type and assay. If you know the concentration range expected in the samples, you can dilute the more concentrated samples to ensure the result falls within the middle range (or linear range) of the standard curve. If the concentration is unknown, we would suggest preparing several dilutions (for example 1:2, 1:10, and 1:100)  to determine a dilution range for your samples.