What are the top categories of enzyme-linked immunosorbent assay (ELISA)?
With the advent of technology, in today’s time, by leveraging ELISA kit, substrates and chromogenic agents are used to generate a change in color that indicates the existence of a specific antigen. Using highly advanced chromogenic agents or reporters, the measurement of antigens in a single assay is possible in an effective manner. ELISA was created as an alternative to radioimmunoassays. ELISA is a plat-oriented assay that recognizes and calculates antigens, proteins, antibodies, hormones, and peptides. The initial step of carrying out the ELISA experiment is to immobilize the specific antigen of a body that remains present within the multiwall plate. Immobilization is done through two primary methods.
Direct ELISA method
In comparison to other methods, the direct ELISA method is much efficient and easy. Other methods have limited steps to follow. An antigen can be directly captured that stays attached on the membrane of the multiwall plate. You can add an enzyme-oriented primary antibody to detect the antigens. In this direct method, chances of error are less due to a few steps that can be efficiently executed by a small number of reagents. Granzyme B ELISA Kit allows you to conduct a direct-ELISA method efficiently. This direct method does not require a secondary antibody that tends to cross-reacting.
Sandwich ELISA method
The sandwich ELISA method makes use of a pair of antibodies. A monoclonal antibody is used for capturing and a polyclonal antibody for the detection of antigens. Leveraging the sandwich ELISA method, you can allow the antibodies to become more specific towards the antigen epitope. Even it is considered to be the most effective method among all ELISA categories that can easily recognize antigen containing two epitopes. The ELISA multiwall plates get coated by a monoclonal antibody. The sandwich method offers high sensitivity in comparison to the direct ELISA method and indirect ELISA method. Due to the involvement of a couple of antibodies for the detection of antigen, a high-specific enzyme reaction occurs.
Indirect ELISA method
Higher sensitivity gets exhibited in the indirect ELISA method because it makes use of an enzyme-labelled secondary antibody that attaches with a primary antibody. Due to the lesser requirement of equipment, the indirect ELISA method is much economical. However, due to the cross-reactivity of secondary antibodies with a specific antigen, background noise can increase. This method also takes a considerable amount of time compared to the direct ELISA method. You can use the Granzyme B ELISA Kit to conduct the indirect ELISA procedure, which aids in calculating the actual concentration of primary and secondary antibodies in given samples.
The categories of enzyme-linked immunosorbent assay (ELISA) mentioned above can help you to gain a comprehensive understanding of the most suitable ELISA methods. You will be able to make use of the best way whenever you require capturing and detecting an antigen using a primary or a secondary antibody.