AlphaLISA & AlphaPLEX

Amplified Luminescent Proximity Homogeneous Assay

AlphaLISA immunoassays are homogenous bead-based luminescent amplification proximity assays that are highly sensitive and offer a large dynamic range and utilize small sample sizes. AlphaLISA is a proximity assay that involves the production of a luminescent/fluorescent signal after energy is transferred between molecule-bound Acceptor and Donor beads. Donor beads include the photosensitizer, phthalocyanine, which transforms ambient oxygen to a reactive form of O2, singlet oxygen, after illumination at 680nm. The excited singlet oxygen has a half-life of 4usec and within that time can diffuse ~200 nm in solution; if an Acceptor bead is present within that space, energy from the singlet oxygen is transmitted to thioxene derivatives in the Acceptor bead. This energy exchange then results in the light production at 615 nm. If there is no Acceptor bead present, the singlet oxygen falls to its ground state and there is no signal.

  • We are able to use various sample types, including serum, plasma, cerebrospinal fluid (CSF), tissue extract, cell culture supernatant, cell lysate, and more. 
  • Targets include enzymes, receptor-ligand interactions, low-affinity interactions, second messengers, DNA, RNA, proteins, protein-protein interactions, peptides, and small molecules.  
  • The assay format can be performed in 96-, 384-, and 1536-well format and used for high throughput screening (HTS).

AlphaLISA can Analyze:

  • Cytokines and biomarkers
  • Cellular epigenetic markers
  • Cellular phosphorylation

Compared to Traditional ELISAs, AlphaLISA Includes:

  • No wash steps
  • Faster results
  • Small sample size (~5uL)
  • Wider dynamic range (up to 5 log)
  • Higher sensitivity
  • Multiplex capabilities
  • Ability to study large protein complexes

For more information about our AlphaLISA platform or the different AlphaLISA kits available, please contact [email protected].


AlphaPLEX is similar to AlphaLISA since it is also a homogenous luminescent proximity assay, but it can analyze multiple analytes in a single well by using multiple Acceptor beads. These analytes can range from small to large proteins in rare biological samples, such as primary or stem cells.

AlphaPLEX Applications:

  • PD/PK biomarkers
  • Stem cell biomarkers
  • Epigenetic markers (total histone vs. specific marker)
  • Amyloid peptides
  • IgG profiling/testing
  • Assay normalization with housekeeping proteins

Our Example Data