Technology

To translate the benefits of CaptSure™ to detection of multiple analytes in individual samples, TGR has expanded its platform with the development of multiple (10) new CaptSure™ anti-peptide antibodies. Each monoclonal CaptSure™ antibody in the CaptSure™ Multiplex system binds a specific peptide that is covalently attached to the primary assay antibody of choice. This allows differential immuno-capture of selected assay targets, providing multiplexing with generic assay system benefits.

TGR’s CaptSure™ ELISA system delivers a quantum-shift in the way an ELISA, the workhorse of many cell-biology labs, can be run. A sample of interest (e.g. cell lysate, serum, conditioned media etc) is simply mixed with assay reagents, incubated in a CaptSure™ microplate, washed and incubated with substrate. The whole process can take as little as an hour, with far fewer handling steps than a conventional ELISA format. Abcam have built a huge catalogue (> 1300) of SimpleStep ELISA products on CaptSure™ making use of their own high quality RabMabs portfolio. Key CaptSure™ ELISA attributes: • Sensitive – for challenging biology • Fast – results in as little as 1 hour • Simple protocol – easy to run • Quantitative – for greater reliability • Flexible – measure multiple analytes on the same plate • Uses most plate readers – colorimetric readout

In association with Revvity, TGR BioSciences’ flagship assay product, AlphaLISA® SureFire® Ultra™ caters primarily for the demanding needs of pharma/biotech in drug discovery. SureFire® is an innovative immunoassay technology platform that addresses complex cell signalling biology. Our extensive offering of > 400 ‘Phospho’ and ‘Total’ protein assays are pivotal to the characterization of various cellular processes and disease states and their reliable and sensitive detection helps to advance both drug discovery and basic research.

AlphaLISA® SureFire® Ultra™ assay kits allow the rapid, sensitive and quantitative detection of phosphoproteins from cells. The kits utilize Alpha Beads that are each coated to specifically capture one of the assay antibodies. The Donor Bead is coated with streptavidin to capture the biotinylated antibody while the Acceptor Bead is coated with a proprietary “CaptSure™” antibody that immobilizes the other antibody which is labeled with a CaptSure™ tag. In the presence of a target protein (i.e. phosphoprotein in schematic below), the two antibodies bring the Donor and Acceptor Beads in close proximity, enabling the generation of an Alpha signal upon illumination of Donor beads by an Alpha-enabled plate reader (see below). The amount of light emission is directly proportional to the quantity of phosphoprotein present in the sample.

SureFire® assays are also highly suited to targeted protein degradation approaches with the emergence of PROTAC (PROteolysis TArgeting Chimera) technology in recent years. This enables researchers to combine the measurement of cellular ‘Total’ protein level changes induced by PROTAC degraders with the functional ‘Phospho’ readouts that SureFire® assays are highly renowned for.

In association with Revvity, TGR more recently launched a multiplex range of products called Multiplex SureFire Ultra™ (MPSU) kits. These MPSU assays allow the rapid, sensitive, and quantitative detection of phosphoproteins from cells, combined with the measurement of the total amount of the same protein in the same well. This Alpha Multiplex measurement is achieved by the use of two Alpha Acceptor Beads that emit at distinct wavelengths (545 nm and 615 nm). The two distinct Alpha Acceptor Beads report their binding to a distinct antigen through their association with specific assay antibodies, as indicated below. Full assay catalog listing is available at: www.revvity.com/category/alpha-surefire-kits The Alpha 615 (Europium) Acceptor bead is conjugated with the CaptSure™ antibody, and the anti-phospho target antibody binding this bead is tagged with the CaptSure™ peptide. The Alpha 545 (Terbium) Acceptor bead is coated with the CaptSure2™ antibody, which binds the CaptSure2 peptide-tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody. The assay utilizes the bead-based Alpha Technology and requires an Alpha Technology-compatible plate reader capable of reading dual emission wavelengths. For more information about the AlphaPlex® technology download the “AlphaPlex Quick Start Guide” and the “AlphaPlex Assay Development Guide” to find guidance about filters and mirrors selection, instrument protocol and channels crosstalk correction.