Biotin coated - Evaluation of binding specificity towards Streptavidin-peroxidase conjugate

  1. Dilute streptavidin-peroxidase conjugate from 1: 500 to 1: 8000 with 0,1 M PBS pH 7,2 containing 0,2 % BSA.
  2. Add 100µl of each dilution to the wells of Biotin coated plate and incubate 60' RT. Add the same solutions to abumin coated plate as comparison for evaluate the specificity of binding.
  3. Leave blank wells as control
  4. Empty the wells and wash with 0,1M PBS pH 7,2 + 0,05% Tween® 20 four times
  5. Add 100 µl /well of TMB substrate solution and incubate 10 minutes at room temperature
  6. Stop the substrate reaction by adding 100 µl of sulphuric acid 1 N and read the optical density values at 450 nm.