What are the important features to check in ELISA kits?
Posted: Nov 18, 2019
ELISA (Enzyme-linked Immunosorbent Assay) kits make it easy for you to detect molecules and perform ligand-binding assays successfully. ELISA kits help in the determination of antigens in unknown samples. It is much better to get the kit than developing your own from scratch. The kits are more accurate, convenient, and faster in their ability to detect antigens. However, there is a broad range of ELISA kits available in the market, and knowing which one to opt for might feel difficult. So, take a look at these two important features that an ELISA kit needs to have.
There are many forms of ELISA kits, and a common character among them is that they can capture the antigen. They can absorb it to the assay plate or capture it with the antibody, which gets attached to the assay plate. You will have to use the right format and container system for the experiment. The procedure is usually done using 384 or 96 plates, but using immune-spot and in-cell formats are also common. The in-cell technique detects the antigens using the antibody in fixed cells, which are cultured in the micro-plates.
Sandwich assay is another common format for ELISA. The name comes from the fact that the antigen stays bound between two antibodies, one for detecting and the other for capturing. The format has many takers because it is robust and sensitive. However, it is not a favorable method if the antigen is small or has only one antibody binding site.
Types of antibodies
Polyclonal and monoclonal antibodies get used in ELISA, and a combination can also prove useful in many cases. However, a couple of points have to be kept in mind when using these antibodies. It is useful to go for a polyclonal antibody for capturing and monoclonal antibody for detection for the sandwich assay. It will enable you to capture all antigens with the polyclonal antibody and detect the specific subsets with the monoclonal antibody.
Detection and capture antibodies need to identify different epitopes on antigens that don't overlap. This fact is important because both the antibodies for Antibody-antibodies Immunoassays must not compete for the same binding site on the antigen. Thus, the detection and capture antibodies need to come together to make a good team because otherwise, the interference between them will lead to skewed assay results. A matched pair of detection-capture antibodies will work the best and lead to no problems. There are antibody suppliers who make sure that this information gets mentioned beforehand.
It will seem easier to choose an ELISA kit when you keep these two factors in mind. However, the most important factor is always whether it can detect what you want to detect.
ELISA offers the advantage of having high specificity and sensitivity. There is high throughput, and it is easy to perform. There is hardly any hands-on time, and the protocol is super-easy to carry out. The fact that it is quantitative and can determine the antigen concentration is another plus.
Fareed is a graduate of computer science engineering, a writer and marketing consultant. he continues to study on Nano technology and its resulting benefits to achieving almost there.