Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Lyophilized Standard
- HRP-Conjugated Trastuzumab
- TMB One-Step Substrate
- Stop Solution
- Assay Diluent
Other Materials Required
- Microplate reader capable of measuring absorbance at 450nm
- 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
- Log-log graph paper or computer and software for ELISA data analysis
- Horizontal orbital microplate shaker
- Tubes to prepare standard and sample dilutions
Protocol Outline
- Prepare all reagents, samples and standards as instructed in the manual.
- Add 100 µl of standard or sample to each well. Incubate 2 hours at room temperature.
- Add 100 µl prepared HRP-conjugated Trastuzumab to each well. Incubate 1 hour at room temperature.
- Add 100 µl TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
- Add 50 µl of 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.