What outcome results from improper washing of a tube or well after adding the enzyme-antibody conjugate in an ELISA system?

What outcome results from improper washing of a tube or well after adding the enzyme-antibody conjugate in an ELISA system?

• A. Result will be falsely decreased
• B. Result will be falsely increased
• C. Result will be unaffected
• D. Result is impossible to determine

Answer: (B)

Improper washing of a tube or well after adding the enzyme-antibody conjugate in an ELISA system can lead to a falsely increased result. This occurs because inadequate washing fails to remove unbound or excess enzyme-antibody conjugate from the surface. When this unbound conjugate is not washed away, it can contribute to higher levels of signal in the assay, resulting in an elevated optical density (OD) reading. As a result, the assay interprets this increased signal as a higher concentration of the target analyte than is actually present in the sample. Accurate washing is crucial in ELISA procedures to ensure that only the specifically bound conjugate remains, which allows for a more accurate reflection of the true concentration of the analyte being measured.

Improper washing of a tube or well after adding the enzyme-antibody conjugate in an ELISA system can lead to a falsely increased result. This occurs because inadequate washing fails to remove unbound or excess enzyme-antibody conjugate from the surface. When this unbound conjugate is not washed away, it can contribute to higher levels of signal in the assay, resulting in an elevated optical density (OD) reading.

As a result, the assay interprets this increased signal as a higher concentration of the target analyte than is actually present in the sample. Accurate washing is crucial in ELISA procedures to ensure that only the specifically bound conjugate remains, which allows for a more accurate reflection of the true concentration of the analyte being measured.