Editing Elisa protocol (section)

== Method ==

Coat microtiter plate wells with 100 µl of the antigen solution, at a concentration of between 1-10 µg/ml in coating buffer. Cover the plate and incubate overnight at 4°C. Wash the plate 3 times in wash buffer.

Add 150 µl of blocking solution to each well. Incubate for 1 hour at 37°C. Wash 4 times in wash buffer.

Add 100 µl of unconjugated detection antibody (appropriately diluted in wash buffer) to each well. Incubate for 1 hour at 37°C. Wash 3 times in wash buffer.

Add 100 µl enzyme-conjugated secondary antibody (appropriately diluted in wash buffer) to each well. Incubate for 1 hour at 37°C. Wash 3 times in wash buffer.

Add 100 µl of the appropriate substrate solution to each well. Incubate at room temperature (and in the dark if required) for 30 minutes, or until desired color change is attained.

Read absorbance values immediately at the appropriate wavelength or add 50 µl of “stop solution”. Gently tap plate to ensure thorough mixing. Measure absorbance within 30 minutes.