Indirect ELISA

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Antibody Detection

An indirect Enzyme-Linked Immunosorbent Assay (iELISA) is a common biochemical technique that detects an antibody using a labeled secondary antibody. This assay is primarily used to detect the total antibody amount in a sample. This method is commonly utilized to diagnose infection and to quantify antibodies against an invading antigen.

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How It Works

RayBio iELISAs employ a specific antigen-coated 96-well plate. A sample containing the primary antibody is incubated into the wells and the antibodies present bind to the immobilized antigen. Next, the wells are washed and a biotin-labeled detection antibody that recognizes the primary antibody is added. After washing away unbound antibodies, an HRP-conjugated secondary antibody is then added, and when combined with a substrate, produces a signal amplification that is directly proportional to the amount of bound sample antibody.

How indirect ELISAs work step-by-step


  1. High sensitivity
    1. Multiple labeled antibodies can bind a single antigen
    2. Maximum primary antibody binding since it is not labeled
  2. Adaptable
    1. Different primary antibodies can be used with a single biotin-labeled detection antibody
    2. Different detection systems can be used with the same primary antibody (colorimetric, chemiluminescent, etc.)
  3. Low cost
    1. Fewer labelled antibodies are needed
  4. Reproducible and reliable
  5. Results within 3 hours

Why RayBiotech ELISA Kits?

  • We manufacture, not resell, ELISA kits. Right here in the USA.
  • Our ELISAs are ISO certified and validated for exceptional reproducibility. Read more about our quality control processes.
  • All RayBio® ELISAs are backed by our 90-day 100% satisfaction guarantee.
  • Discounts and incentives available for publications citing our products, and for product reviews.