ELISA Methodology

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(Name) (Instructors’ name) (Course) (Date) ELISA Protocol ELISA (Enzyme-linked immunosorbent assay) is an assay procedure which is created for checking and quantifying substances like the proteins, peptides, hormones, and antibodies. There are many forms of ELISA which make the ELISA protocol to have few variations. The different types of ELISA protocol include; sandwich, direct, elispot, indirect and competitive ELISA protocol. However, the major ELISA protocol and process are similar. All in all, the ELISA protocol involves three majors steps; plate preparation, assay protocol, and analysis of results. Plate Preparation This process involves diluting the extracted antibody to the working samples in the plates. Afterward, you should cover a 96-well microplate with 150 µL per plate of the captured body which is diluted. You should then enhance the sealing of the plate and incubate it overnight at 4 ºC. Aspiration follows which involves washing the plate with at least 300 µl wash buffer and replicate the procedure twice through several washes. To achieve good results, it is crucial to complete the extraction of fluid at every step. After the final wash, you should dispose any residual wash by overturning the plate and use a paper towel for blotting purposes. The last step involves blocking the plate by including 300 µL of blocked buffer to each plate. Assay Process The assay process involves adding 150 µl of standards or sample in the dilute sample solution on each plate. Enhance sealing of the plates and incubating for 2 hours at normal temperature and repeat step 2 of plate preparation that is the aspiration or washing. Add 150 µL of the antibody

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