ELISA Development

ELISA Protocol

Quantitative detection of antigens using LabReCon ELISA kits.

Sandwich ELISA Protocol

1. Coat plate with capture antibody (100 μL/well) in coating buffer, overnight at 4°C
2. Wash 3× with PBST
3. Block with 1% BSA/PBS, 1 hour at RT
4. Wash 3× with PBST
5. Add standards and samples (100 μL/well), incubate 2 hours at RT
6. Wash 5× with PBST
7. Add detection antibody (100 μL/well), incubate 1 hour at RT
8. Wash 5× with PBST
9. Add streptavidin-HRP (100 μL/well), incubate 30 min
10. Wash 5× with PBST
11. Add TMB substrate (100 μL/well), incubate 15 min in dark
12. Stop with 2N H₂SO₄ (50 μL/well)
13. Read at 450 nm within 30 min