ELISA, short for enzyme-linked immunosorbent assay, is a routine method performed in molecular biology and life sciences labs for quantitative analysis of a range of molecules. It includes hormones, peptides, proteins, and some other small molecules in a cell lysate or other samples.
Other than these, the assay is also used to detect growth factors, cytokines, chemokines, and SARS-CoV-2-related proteins. The ELISA assay also often goes by enzyme immunoassay (EIA).
The four common types of ELISA assay include:
This assay has high sensitivity and is less expensive compared to other ELISA assays.
Figure: An illustration of the four types of ELISA assay.
In this article, we will cover more about the steps involved in ELISA assay and the reagents required to perform this high-throughput assay in labs. Further, we will also learn the types of ELISA kits available today in the market to make your lab life easier.
Although different ELISA formats are available today to assist in lab workflows, they all have these common steps:
Figure: An illustration of ELISA assay.
Scientists perform ELISA either by creating their own ELISA kit using individual reagents, based on their workflow or research goals, or by purchasing commercially available ELISA assay kits. The kits are either available based on the types of ELISA plates or ELISA formats, such as sandwich ELISA kit and competitive ELISA kit.
In addition to reliability testing, some manufacturers also test the kits for fit-for-purpose validation and stability. This ensures their precision, sensitivity, accuracy, specificity, and reproducibility for the assay.
The kit includes ELISA plates precoated with an antigen-specific antibody. It also contains a biotinylated detection antibody, recombinant protein standard, and buffers and reagents for colorimetric detection. The kit is fast, easy to use, and tested by companies for validation and plate variability.
The kit contains reagents and materials to coat your own ELISA plates and run the assay. It’s suitable if you have a limited budget in hand and are not concerned with inter- and intra-assay variance. It’s flexible, affordable, and optimized for different ELISA assays.
Some other ELISA development kits available in the market include Chemiluminescent ELISA Kits which are used to detect key biomarkers during research and ProQuantum Kits which are high-sensitivity kits and work on proximity-based amplification technology.
ELISA kits are used for the detection and quantification of a range of molecules in different sample types, such as:
Though some scientists prefer to prepare their own reagents and materials to perform ELISA assay, some labs prefer the kits because they are:
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ELISA is a biochemical technique used in life sciences labs to detect and quantify a range of molecules, including peptides, proteins, and hormones. To make the workflow faster and more convenient for labs, a spectrum of ELISA kits are available in the market.
You can either find them based on the ELISA formats, such as sandwich ELISA format and competitive ELISA format, or types of molecules you need to study, such as Protein A ELISA Kit, Human ICAM4 PicoKine ELISA Kit, and Human CD51 PicoKine ELISA Kit. Other than these, ELISA kits based on the types of microplates are also available, which include Precoated Instant Kits and Unocated Antibody Pair Kits.
It is crucial to use high-quality kits and chemicals for performing assays that have been thoroughly tested for their performance. Additionally, high-throughput instruments should be used in combination with these reagents to ensure the acquisition of reliable and validated assay outcomes.
If you are a lab just starting and have limited financial resources, the cost of conducting certain assays can be prohibitively expensive. This can stall your research progress and make it challenging to achieve your desired outcomes.
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