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AlphaLISA is a no-wash ELISA alternative and can be used for applications such as:
In a typical AlphaLISA assay, 1 mg of Acceptor beads is sufficient to run 1,000-2,000 wells using a 50 µL reaction volume.
|Bead Type or Core Bead Type||AlphaLISA Acceptor|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5 mg|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
Biological drug products often elicit an immune response in patients that can lead to clinical consequences of the presence of anti-drug antibodies (ADA) varying from mild to serious adverse events. Therefore, the presence of ADA is a major safety and efficacy concern and should be evaluated and correlated with any pharmacological or toxicological observations.
The development of rapid and sensitive assay platforms for ADA detection is an essential step of the drug development process. In this application note, we demonstrate that the AlphaLISA® mix-and-read homogeneous assay permits the sensitive detection of ADA.
Alpha has been used to study a wide variety of interactions, including protein:protein, protein:peptide, protein:DNA, protein:RNA, protein:carbohydrate, protein:small molecule, receptor:ligand, and nuclear receptor:ligand interactions. Both cell-based and biochemical interactions have been monitored, and applications such as phage display, ELISA, and EMSA (electrophoretic mobility shift assay) have been adapted to Alpha.
This guide presents the simple conversion of an ELISA or other immunoassay to an AlphaLISA® immunoassay.
AlphaScreen® and AlphaLISA® are bead-based assay technologies used to study biomolecular interactions in a microplate format. The acronym “Alpha” stands for Amplified Luminescent Proximity Homogeneous Assay. The assay does not require any washing steps. Binding of proteins or other binding partners captured on the beads leads to an energy transfer from one bead to the other, ultimately producing a luminescent signal.
Binding events between biomolecules are important components of biological processes and a number of these biomolecular interactions have been targeted for the development of novel therapeutic drugs. p53 is a transcription factor and tumor suppressor protein that is activated in response to cellular stress, and MDM2 was identified as a negative regulator that binds to p53 and tags it for ubiquitination and subsequent degradation.
The p53-MDM2 protein-protein interaction has been an excellent target for therapeutic drugs and therefore makes a good model system for developing an AlphaLISA assay to screen for inhibitors of the interaction. In this technical note, we show how to develop an assay to screen for inhibitors and how to measure a dissociation constant for moderate binding protein-protein interaction using AlphaLISA®.