1.0 Dilute reference and investigated samples.
- Dispense 100μl of prepared dilutions of investigated and reference samples per well.
- Incubate 90 minutes at 37 °C, shake gently.
2.0 Prepare biotinylated anti-Rabies antibody.
- Wash plates 2x time.
- Dispense 100μl of biotinylated anti-Rabies antibody diluted to 1/100 to all wells.
- Incubate 30 minutes at 37°C, shake gently.
3.0 Prepare streptavidin peroxidase conjugate.
- Wash plates 4x time.
- Dispense 100μl of streptavidin peroxidase conjugate diluted to 1/100 to all wells.
- Incubate 30 minutes at 37°C, shake gently.
4.0 Wash plates 4x time.
- Dispense 100μl of TMB Substrate to all wells.
- Incubate 15-20 minutes at room temperature (18 – 25°C), shake gently.
5.0 Stop reaction with 50μl of Stop solution.
- Read absorbance (OD) at 450 nm.
- Check validation criteria.
- Interpret result.
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