Remove NDV coated plate from sealed bag and record location of samples on template.
2. Add 100 ml of negative control into wells A1 and B1.
3. Add 100 ml of positive control into wells C1 and D1.
4. Add 100 ml of diluted samples into all the appropriate wells. Cover plate with lid and incubate at
room temperature (22-27°C) for 30 minutes.
5. Aspirate contents of wells and wash 4 times with wash buffer (350ml per well). Invert plate and tap
firmly on absorbent paper until no moisture is visible.
6. Add 100 ml of Conjugate reagent into all the appropriate wells. Cover plate with lid and incubate
at room temperature (22-27°C) for 30 minutes.
7. Aspirate contents of wells and wash 4 times with wash buffer (350ml per well). Invert plate and tap
firmly on absorbent paper until no moisture is visible.
8. Add 100 ml of Substrate reagent into all the appropriate wells. Cover plate with lid and incubate at
room temperature (22-27°C) for 15 minutes.
9. Add 100 ml of Stop solution to appropriate wells to stop reaction.
10. Blank the microtitre plate reader on air and record the absorbance of controls and the samples by
reading at 405 nm.