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There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common types of ELISA is the so-called "sandwich ELISA." It is termed this because the antibody that you are detecting gets sandwiched between an antigen and a chromogenically-conjugated antibody. Below you will find a basic protocol for this assay.
*Common Substrates and the appropriate plate reader setting
- ABTS: 405-410 nm
- TMB: non-stopped 620-650 nm, stopped 450 nm
- OPD: non-stopped 450 nm, stopped 490 nm
- pNPP: 405-410 nm
- BluePhosTM: 595-650 nm