Kit Components
| Component |
Size / Description |
| Pre-Coated 96-Well Microplate |
1 plate |
| Wash Buffer |
1 vial |
| Assay Diluent |
1 vial |
| Ligand Protein |
2 vials |
| Biotinylated Detection Antibody |
2 vials |
| HRP-Streptavidin |
1 vial |
| TMB One-Step Substrate Reagent |
1 vial |
| Stop Solution |
1 vial |
Additional Materials Required
- Microplate reader capable of measuring absorbance at 450 nm
- Shaker
- Pipettes capable of accurately delivering 2 µL to 1 mL volumes
- Pipettes capable of delivering 1-25 mL volumes for reagent preparation
- 100 mL and 1 liter graduated cylinders
- Distilled or deionized water
- Tubes to prepare sample dilutions
Assay Procedure Summary
- Prepare all reagents and Test Reagent samples as instructed.
- Add 100 µL Test Reagent samples to each well. Incubate 2.5 hours at room temperature or overnight at 4°C.
- Add 100 µL the prepared Biotinylated Detection Antibody solution to each well. Incubate 1 hour.
- Add 100 µL the prepared HRP-Streptavidin Concentrate solution to each well. Incubate 1 hour.
- Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 min.
- Add 50 µL Stop Solution directly to each well.
- Read at 450 nm immediately.
Storage/Stability
The kit may be stored at 4°C up to 1 month from the date of shipment. Shelf life can be extended up to 6 months by storing protein and antibodies, at -20°C immediately upon delivery. All other kit components should be stored at 4°C.