UbiQuant S ELISA and AlphaLISA: High-Throughput Cellular Ubiquitylation Assays
LifeSensors’ UbiQuant S ELISA and AlphaLISA assays are industry-leading platforms for measuring ubiquitylation of a target protein in cells. These assays utilized TUBEs (Tandem Ubiquitin Binding Entities) to detect ubiquitylated substrates, and are individually optimized for a specific substrate protein.
UbiQuant S ELISA and AlphaLISA are both a service and a kit.
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Conceptual Schematic of the UbiQuant S ELISA Assay
Polyubiquitlyated proteins are captured by an immobilized ubiquitin affinity matrix. Subsequently, unbound proteins are washed away, and ubiquitylated protein of interest bound to the matrix is identified using a POI-specific antibody and traditional ELISA detection reagents.
Conceptual Schematic of the UbiQuant S AlphaLISA Assay
TUBEs conjugated to AlphaLISA donor beads bind to polyubiquitylated proteins from a cell lysate. Simulataneously, POI antibodies conjugated to AlphaLISA acceptor beads bind to the POI in the cell lysate. If ubiquitylated POI is present, the concurrent binding of antibody and TUBE brings the AlphaLISA donor and acceptor beads into close proximity, resulting in a luminescent signal that is quantitatively correlated to the level of ubiquitlyated POI.
Sample Data From UbiQuant S ELISA and AlphaLISA Assays
Jurkat cells were treated with bortezomib (BTZ) or BTZ in combination with selective USP7 inhibitor P0217564 (564). Following treatment and cell lysis, Ubiquitylation of USP7 was measured by the UbiQuant S ELISA and AlphaLISA assays.