Reagents and Materials:

Biocarbonate buffer- Prepare the 50mM Biocarbonate buffer pH 9.2 in distilled water and filter sterile with 0.2µm filter.

ELISA plate: Flat Bottom 96 well is more suitable for performing ELISA.

Antigen solution: Prepare 5µg/ml antigen solution in biocarbonate buffer pH 9.2.

BSA: Prepare 10mg/ml BSA solution in distilled water and filter sterile with 0.2µm filter.

Primary antibody and secondary antibody

PBS containing Tween 20

Procedures: Different steps in performing in-direct ELISA is given in the Figure 35.2. These steps are as follows:

Figure 35.2: Different Steps in performing in-direct ELISA to measure antibody level in the serum.