How to perform competitive ELISA?

1.ELISA plates were coated with 100 μl of the mutant peptide (200 ng/ml) overnight.
2.After blocking plates with PBS with 1% BSA for 1 h, the peptide-specific monoclonal antibodies (10 ng/ml) pre-incubated overnight with varying concentrations (0–1,000 μg/ml) of native peptides in PBS with 1% BSA was added to the mutant peptide-coated plates, and we performed ELISA as described above.
3.For the alanine scanning mutagenesis experiments, competitive ELISA was performed as described above using peptide variants.