Since its inception in 1971, the ELISA method has become one of the greatest tools researchers have to generate robust, high throughput data. Originally, it was developed as a chromogenic replacement for radioimmunoassays. As with its predecessor, this technique exploits the basic immunology concept that an antigen (protein, peptide or hormone), found in a fluid sample binds to a specific antibody. The antibody bound to the target can either be directly conjugated to an enzyme or incubated with a secondary antibody-enzyme conjugate. Upon incubation with a substrate, the reaction produces a signal that can be measured with proper instrumentation.
The ELISA is a very flexible method, allowing for quantitation of an antigen or antibody in a sample. The ELISA method is most commonly used to determine serum antibody concentration, such as in the HIV test as well as some allergic diseases such as food allergies. There is constant pressure to develop assays with increased sensitivity for early disease detection. Accurate and robust information is paramount in guiding therapeutic decision making for optimal patient care.
There are four different ELISA formats (direct, indirect, sandwich and competitive) to accommodate sample type, available reagents and desired sensitivity. Each of these formats has its own advantages and disadvantages. However, all the various formats can be easily upgraded by making the switch from chromogenic to chemiluminescent detection. Chemiluminescent detection offers higher sensitivity, lower background and a wider dynamic range. Advansta’s ELISABright is an enhanced and highly sensitive chemiluminescent HRP substrate optimized for chemiluminescent ELISA applications. This luminol-based substrate demonstrates a wide linear dynamic range and a superior signal to noise ratio for excellent quantitative ability. ELISABright allows for enhanced detection of low abundance proteins and reduced consumption of antibodies.
To make the switch from chromogenic to chemiluminescent detection you will need to invest in new instrumentation. One of our favorite chemiluminescent plate readers is Perkin Elmer’s EnVision® Multimode Plate Reader. Another option includes the SpectraMax® M5 Plate Reader from Molecular Devices. Whichever plate reader you decide on, chemiluminescent detection will be an investment that is well worth the upgrade.
ELISABright Chemiluminescent Substrate
AdvanBlock EIA Blocking Solution
HRP Conjugated Secondary Antibodies
10X EIA Coating Buffer