ELISA, western blot, and other immunodetection technologies have commonly been used for quantitation of biomarkers and other analytes in a variety of sample types. These methods necessitate long, tedious protocols and often provide limited detectable range.
AlphaLISA® provides a homogenous (no-wash) alternative to ELISAs, with a simple, streamlined workflow that can be used to detect and quantitate biomolecules in both simple and complex sample types. AlphaLISA is a bead-based luminescent amplification assay, offering greater sensitivity, a wider dynamic range and small sample sizes over traditional ELISA.
In the AlphaLISA assay, a biotinylated antibody and an antibody-conjugated AlphaLISA Acceptor bead are used to capture the target analyte. The biotinylated antibody associates with an Alpha streptavidin-coated Donor bead. When the analyte is present in the sample, the Donor and Acceptor beads are brought together. Upon excitation, a photosensitizer inside the Donor bead converts ambient oxygen to an excited singlet state. Singlet oxygen diffuses up to 200 nm to produce a chemiluminescent reaction in the Acceptor bead, leading to light emission. The amount of light is proportional to the amount of analyte present in your sample.
Alpha offers the benefits over ELISAs including:
- No-wash
- Faster time to results
- Requires a fraction of the sample size, ~5µL
- Wider dynamic range up to 5 log
- Improved sensitivity
- Ability to multiplex
- Ability to study large protein complexes
For research use only. Not for use in diagnostic procedures.