PerkinElmer

Alpha SureFire Kits

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Alpha SureFire® Technology is a reagent platform for rapid, sensitive, and quantitative detection of protein phosphorylation from cells. It is based on the proven Alpha Technology, a very versatile platform that eliminates time-consuming separation and wash-steps. The SureFire sandwich bead-based immunoassays bring Alpha Donor and Acceptor beads together, causing a cascade of chemical reactions to create a greatly amplified signal to measure endogenous protein phosphorylation. It can also be used with transfected cells and enables the precise interrogation of various signalling pathways, receptors and kinase targets in a cell-based format.

All the Alpha SureFire kits provide:

  • All-in-one-well alternative to Western Blotting
  • Flexibility to use cultured cell lines, stem cells and primary cells
  • Cost-effective and flexible – a single plate can accommodate any assay for any target
  • Great sensitivity and dynamic range
  • Easy automation
  • No harmful chemicals and non-radioactive, eliminating special reagent handling and disposal

Alpha SureFire Multiplex assay principle
Alpha SureFire Multiplex assay principle

Alpha SureFire® Ultra Multiplex

The Alpha SureFire® Ultra™ Multiplex assay kits allow the rapid, sensitive, and quantitative detection of phosphoproteins from cells, combined with the measurement of the total amount of the AKT, ERK or cofilin protein or combined with the measurement of a second phosphoprotein. This Alpha Multiplex measurement is carried out in the same assay plate well from a single sample of cell lysate, and is achieved by the use of two types of Alpha Acceptor beads that emit at distinct wavelengths (545nm and 615nm). The two distinct Alpha Acceptor beads report their binding to a distinct antigen through their association with specific assay antibodies, as indicated below.

Alpha SureFire Ultra Multiplex assays are divided into two categories (1) Multiplex SureFire Ultra (MPSU) kits and (2) Terbium SureFire Ultra (TBSU) kits. The MPSU kits are complete kits for phospho and total detection of the same target (ERK or AKT), while the TBSU kits measure a single phosphoprotein target and are intended to be combined with a standard AlphaLISA SureFire Ultra kit (ALSU) to assess the second target. The TBSU kits allow for a pick-and-pack combination of any two phosphoprotein targets in the assay lists, providing measurement of two separate targets with ultimate flexibility of selection.

  • High sensitivity and dynamic range – detect low levels of protein phosphorylation, with internal normalization for total protein content
  • No interference from spiked antibodies – uses CaptSure™ technology instead of protein A
  • 2- incubation protocol that can be run in 2 hrs for faster results
  • Convenience of choosing HTS-compatible version or 96-well High Volume (HV) format for low-throughput applications - for all the targets
  • More than 1000 combinations of either phospho X phospho target or phospho x total targets are available
AlphaLISA SureFire assay principle
AlphaLISA SureFire assay principle

AlphaLISA® SureFire® Ultra

The next-generation technology for detecting protein phosphorylation, AlphaLISA SureFire Ultra is a highly sensitive assay for screening therapeutic antibodies and small molecules on cultured cells or tissue lysates. AlphaLISA SureFire Ultra combines CaptSure™ technology with AlphaLISA Acceptor beads for a more efficient, faster, and simpler assay protocol.

  • High sensitivity and dynamic range – detect low levels of protein phosphorylation
  • No interference from spiked antibodies – uses CaptSure™ technology instead of protein A
  • Almost 2-fold increase in 384-well plates working volumes for easier pipetting, compared to AlphaScreen SureFire
  • 1- or 2- incubation protocol that can be run in 2 hrs for faster results
  • Convenience of choosing HTS-compatible version or 96-well High Volume (HV) format for low-throughput applications - for all the targets
  • Ideal tool when working with samples such as tissue lysates - AlphaLISA Acceptor beads have better spectral characteristics with regard to hemoglobin light quenching

With CaptSure™ technology, there is no interference from spiked antibodies, and with a lower background signal, you have better sensitivity and dynamic range, resulting in more – and more accurate – information from all your samples.

AlphaScreen SureFire assay principle
AlphaScreen SureFire assay principle

AlphaScreen® SureFire®

 

The traditional platform for a wide variety of applications, AlphaScreen® SureFire® is a no-wash assay for the detection of phosphorylated proteins in cells. The AlphaScreen SureFire kits use an immunosandwich assay based on a combination of protein A and streptavidin for the capture of the sandwich antibodies.

  • Simple protocol that can be run in 4 hrs
  • Great sensitivity and dynamic range
  • Convenience of choosing HTS-compatible version or 96-well High Volume (HV) format for low-throughput applications - for some targets only
  • Currently the widest choice of phospho-targets in our catalogue
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