Application Notes
Kit Components
- Pre-coated 96-well microplate
- Wash Buffer
- Positive Control
- Biotinylated Detection Antibody
- Biotinylated Anti-Positive Control
- HRP-Streptavidin concentrate
- TMB One-Step Substrate
- Stop Solution
- Assay Diluent
- 5X Sample Diluent
Other Materials Required
- Microplate reader capable of measuring absorbance at 450 nm
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Absorbent paper
- Distilled or deionized water
Protocol Outline
- Prepare all reagents, samples and standards as instructed.
- Add 100 µl positive control or sample to each well. Incubate 1 hour at room temperature.
- Add 100 µl prepared Biotinylated detection antibody and Biotinylated Anti-Postive Control into respective wells. Incubate 1 hour at room temperature.
- Add 100 µl prepared HRP-Streptavidin solution to each well. Incubate 30 minutes at room temperature.
- Add 100 µl TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
- Add 50 µl Stop Solution to each well. Read at 450 nm immediately.
Storage/Stability
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.