Determination of human malondialdehyde (MDA) in saliva using an ELISA kit (prepared by Prof. OS)
Saliva samples were centrifuged for 10 min at 4000 xg. The supernatant was separated and used for determination of MDA levels using ELISA Kit Cat No. MBS263626 provided by My BioSource (USA, NY). This kit employs the “Double Antibody Sandwich” technique. The principle of double antibody sandwich is based on the characteristics of a target analytic with more than two possible epitopes that can be identified by both the precoated capture antibody and the detection antibody simultaneously.In this kit, the precoated antibody is an anti-human MDA monoclonal antibody, while the detection antibody is a biotinylated polyclonal antibody. Samples and biotinylated antibodies are added into ELISA plate wells and washed out with PBS or TBS after their respective additions to the wells. Then, avidin-peroxidase conjugates were added to the wells. TMB substrate is used for colouration after the enzyme conjugate has already been thoroughly washed out of the wells by PBS or TBS. TMB reacts to form a blue product from the peroxidase activity, and finally turns yellow after addition of the stop solution (Color Reagent C). The color intensity and quantity of target analytics in the sample are positively correlated [