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Fast immunoassay methods and methods
Rapid immunoassays take advantage of the fact that antibodies are unable to bind to the hydrophobic surface of an unwetted Immobilon®-P transfer membrane. However, they can effectively bind to proteins that have been immobilized on the membrane. These assays are compatible with both chromogenic and chemiluminescent substrates, making them versatile for different detection needs.
Tip: If you're looking to quickly detect high-abundance proteins, a rapid immunoassay is ideal. For more sensitive detection, especially when working with low-abundance targets, standard immunoassays are recommended.
One major benefit of the rapid immunoassay method is that it eliminates the need for blocking steps, which saves time and reduces the risk of errors or inconsistencies (Mansfield, 1994). Additionally, since excess antibody doesn't bind to the dried membrane, fewer washing steps are required. As a result, the entire process can be completed in under two hours, compared to over four hours for traditional methods.
Remember: It's crucial to ensure that the blot is completely dry before beginning a rapid immunoassay. This step is essential to maintain the integrity of the assay and avoid any issues with non-specific binding or poor signal quality.
This streamlined approach makes rapid immunoassays particularly useful in high-throughput settings or when quick results are needed without compromising accuracy. Whether you're a seasoned researcher or just starting out, understanding the differences between rapid and standard immunoassays can help you choose the best method for your specific experimental goals.